CN111417630B - Modulator of Stimulator of Interferon Genes (STING) - Google Patents

Abstract

The present application discloses compounds having the following formula (I): wherein q, r, s, A, B, C, R ^A1 , R A2 , R ^B1 , R ^B2 , R ^C1 , R ^{C2 ,} R ³ , R ⁴ , R ⁵ , R ⁶ , R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ^x and R ^y are as defined herein, or tautomers thereof, or salts thereof, especially pharmaceutically acceptable salts thereof.

Description

Regulators of stimulator of interferon genes (STING)

Related Applications

This application claims priority to U.S. Provisional Application No. 62/568,420, filed on October 5, 2017, the entire contents of which are incorporated herein by reference.

Field of the Invention

The present invention relates to heterocyclic amides useful as modulators of transmembrane protein 173 ((TMEM173), also known as STING (stimulator of interferon genes)) and methods of making and using the same.

Background of the Invention

Vertebrates are constantly threatened by microbial invasion and have evolved immune defense mechanisms to eliminate infectious pathogens. In mammals, the immune system consists of two branches; innate immunity and adaptive immunity. The innate immune system is the first line of defense triggered by pattern recognition receptors (PRRs), which detect ligands from pathogens as well as damage-associated molecular patterns ( Takeuchi O. et al., Cell, 2010: 140, 805-820 ). More and more of these receptors have been identified, including Toll-like receptors (TLRs), C-type lectin receptors, retinoic acid-induced gene I (RIG-I)-like receptors and NOD-like receptors (NLRs) and double-stranded DNA sensors. Activation of PRRs leads to upregulation of genes involved in inflammatory responses, including type 1 interferons, proinflammatory cytokines and chemokines that inhibit pathogen replication and promote adaptive immunity.

The adaptor protein STING (stimulator of interferon genes), also known as TMEM 173, MPYS, MITA and ERIS, has been identified as a central signal transduction molecule in the innate immune response to cytoplasmic nucleic acids ( Ishikawa H and Barber GN, Nature, 2008: 455, 674-678; WO2013/1666000). Activation of STING causes upregulation of the IRF3 and NFκB pathways, leading to the induction of interferon-β and other cytokines. STING is critical for responding to cytoplasmic DNA of pathogen or host origin and to unusual nucleic acids called cyclic dinucleotides (CDNs).

CDNs were first identified as bacterial second messengers responsible for controlling many responses in prokaryotic cells. Bacterial CDNs, such as c-di-GMP, are symmetrical molecules characterized by two 3',5' phosphodiester bonds.

Direct activation of STING by bacterial CDN has recently been confirmed by X-ray crystallography ( Burdette DL and Vance RE, Nature Immunology, 2013: 14, 19-26 ). Therefore, bacterial CDN and its analogs have attracted interest as potential vaccine adjuvants ( Libanova R. et al., Microbial Biotechnology 2012: 5, 168-176; WO2007/054279, WO2005/087238).

Recently, the response to cytoplasmic DNA has been elucidated and shown to be involved in the production of a novel mammalian CDN signaling molecule, called cGAMP, through an enzyme called cyclic GMP-AMP synthase (cGAS, formerly known as C6orf150 or MB21D1), which then activates STING. Unlike bacterial CDNs, cGAMP is an asymmetric molecule characterized by its mixed 2', 5' and 3', 5' phosphodiester bonds. (Gao P et al., Cell, 2013: 153, 1094-1107) . The interaction of cGAMP (II) with STING has also been demonstrated by X-ray crystallography ( Cai X et al., Molecular Cell, 2014: 54, 289-296 ) .

Interferon was first described as a substance that can protect cells from viral infection ( Isaacs & Lindemann, J. Virus Interference. Proc. R. Soc. Lon. Ser. B. Biol. Sci. 1957: 147, 258-267 ) . In humans, type I interferon is a family of related proteins encoded by genes on chromosome 9 that encode at least 13 isoforms of interferon α (IFNα) and one isoform of interferon β (IFNβ). Recombinant IFNα is the first approved biological therapeutic agent and has become an important therapy for viral infection and cancer. In addition to its direct antiviral activity on cells, interferon is known to be an effective regulator of immune responses acting on cells of the immune system.

Administration of small molecule compounds that can modulate the innate immune response, including activation or inhibition of type I interferon production and other cytokines, could become an important strategy for treating or preventing human diseases, including viral infections and autoimmune diseases. This type of immunomodulatory strategy has the potential to identify compounds that are useful not only for innate immunity to infectious diseases, but also for cancer (Zitvogel, L., et al., Nature Reviews Immunology, 2015 15 (7), p405-414) , allergic diseases (Moisan J. et al., Am. J. Physiol. Lung Cell Mol. Physiol., 2006: 290, L987-995) , and neurodegenerative diseases such as amyotrophic lateral sclerosis and multiple sclerosis ( Lemos, H. et al., J. Immunol., 2014: 192(12), 5571-8; Cirulli, E. et al., Science, 2015: 347(6229), 1436-41; Freischmidt, A., et al., Nat. Neurosci., 18 (5), 631-6). , other inflammatory conditions such as irritable bowel disease ( Rakoff-Nahoum S., Cell., 2004, 23, 118(2): 229-41 ), and as a vaccine adjuvant ( Persing et al. Trends Microbiol. 2002: 10(10 Suppl), S32-7 and Dubensky et al., Therapeutic Advances in Vaccines, published online on September 5, 2013 ).

STING is essential for antimicrobial host defense, including protection against a range of DNA and RNA viruses and bacteria (reviewed in Barber et al. Nat. Rev. Immunol. 2015: 15(2): 87-103, Ma and Damania, Cell Host & Microbe, 2016: 19(2) 150-158 ). Herpesviridae, Flaviviridae, Coronaviridae, Papillomaviridae, Adenoviridae, Hepadnaviridae, Ortho- and Paramyxoviridae, and Rhabdoviridae have evolved mechanisms to inhibit STING-mediated type I IFN production and evade host immune control ( Holm et al., Nat Comm. 2016: 7:10680; Ma et al. , PNAS 2015: 112(31) E4306-E4315; Wu et al., Cell Host Microbe 2015: 18(3) 333-44; Liu et al., J Virol 2016: 90(20) 9406-19; Chen et al., Protein Cell 2014: 5 (5) 369-81; Lau et al., Science 2013: 350(6260) 568-71; Ding et al., J Hepatol 2013: 59(1) 52-8; Nitta et al., Hepatology 2013 57(1) 46-58; Sun et al., PloS One 2012: 7(2) e30802; Aguirre et al., PloS Pathog 2012: 8(10) e1002934; Ishikawa et al., Nature 2009: 461(7265) 788-92 ). Therefore, small molecule activation of STING may be beneficial for the treatment of these infectious diseases.

In contrast, increased and prolonged type I IFN production has been associated with a variety of chronic infections, including mycobacteria (Collins et al., Cell Host Microbe 2015: 17(6) 820-8); Wassermann et al., Cell Host Microbe 2015: 17(6) 799-810; Watson et al., Cell Host Microbe 2015: 17(6) 811-9 ), Francisella ( Storek et al., J Immunol. 2015: 194(7) 3236-45; Jin et al., J Immunol. 2011: 187(5) 2595-601 ), Chlamydia ( Prantner et al., J Immunol 2010: 184(5) 2551-60 ), Plasmodium ( Sharma et al., Immunity 2011: 35(2)). 194-207 ) and HIV ( Herzner et al., Nat Immunol 2015 16 (10) 1025-33; Gao et al., Science 2013: 341 (6148) 903-6) . Similarly, excessive type I interferon production has been found in patients with complex forms of autoimmune diseases. Genetic evidence in humans and support from animal model studies support the hypothesis that inhibition of STING leads to a reduction in type I interferon that drives autoimmune diseases ( Crow YJ, et al. , Nat. Genet. 2006; 38 (8) 38917-920 , Stetson DB, et al., Cell 2008; 134 587-598 ). Therefore, inhibitors of STING provide treatment for patients with chronic type I interferon and proinflammatory cytokine production associated with infection or complex autoimmune diseases. Allergic diseases are associated with a Th2-biased immune response to allergens. Th2 responses are associated with elevated IgE levels, which promote hypersensitivity to allergens via actions on mast cells, leading to symptoms such as those seen in allergic rhinitis and asthma. In healthy individuals, the immune response to allergens is more balanced with a mixed Th2/Th1 and regulatory T cell response. Induction of type 1 interferons has been shown to result in a reduction in Th2-type cytokines in the local environment and promote Th1/Treg responses. In this context, induction of type 1 interferons by, for example, activation of STING may provide benefits in the treatment of allergic diseases such as asthma and allergic rhinitis ( Huber JP et al., J Immunol 2010: 185, 813-817).

Compounds that bind to STING and act as agonists have been shown to induce type 1 interferon and other cytokines when incubated with human PBMCs. Compounds that induce human interferon can be used to treat various conditions, such as allergic diseases and other inflammatory conditions such as allergic rhinitis and asthma, infectious diseases, neurodegenerative diseases, precancerous syndromes and cancer, and can also be used as immunogenic compositions or vaccine adjuvants. Recent studies have shown that compounds that bind to STING can act as antagonists and can be used to treat inflammation, such as autoimmune diseases, metabolic diseases, neuroinflammation, and inflammation in the heart that leads to heart disease (e.g., myocardial infarction) ( Ridker et al., N ENG J Med 2017, 377 (12), 1119-1131 ; King et al., Nat Med. 2017 Dec; 23 (12): 1481-1487 ).

Based on recent studies, it is believed that inhibition of cGas or STING may be useful for treating or preventing metabolic diseases such as insulin resistance, non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH), obesity, diabetes, hypertension, fatty liver, and cardiovascular disease ( Qiao et al., Metabolism Clinical and Experimental (2007), 81, 13 – 24; Bai et al., PNAS (2017), 114, no. 46, 12196-12201; Iracheta et al., Journal of Biological Chemistry (2016) 52, 26794-26805; Cruz et al., Molecular Metabolism (2018) 1-11; Patrasek et al., Proc Natl Acad Sci (2013), 110(41): 16544-9; Mao et al., Arterioscler Thromb Vasc Biol. (2017) 37(5): 920-929 ).

It is envisioned that targeting STING with activation or inhibitors may be a promising approach for treating diseases and conditions in which modulation of the type 1 IFN pathway is beneficial and as an immunogenic or vaccine adjuvant, including inflammatory, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, tumor metastasis, metabolic diseases, cardiovascular disease.

Skin cancer and various skin viral infections involve an immunoprivileged environment, and activation of local immune responses to lesions can be a local treatment approach. STING agonists can be used to treat viral warts, superficial skin cancers, and premalignant actinic keratoses. Through a dual mechanism of action, STING activation (e.g., via microneedle patch delivery or topical formulations) can be used to directly control HPV via antiviral type I interferon production and indirectly control HPV by enhancing the adaptive immune response downstream of innate immune activation. STING agonists can activate innate immune responses in lesions and drive anti-HPV T-cell responses.

Recent evidence has shown that spontaneous activation of the STING pathway within tumor-resident dendritic cells leads to type I IFN production and adaptive immune responses against tumors. Furthermore, activation of this pathway in antigen presenting cells (APCs) within the tumor microenvironment drives subsequent T-cell priming against tumor-associated antigens. Corrales and Gajewski, Clin Cancer Res ; 21(21); 4774-9, 2015.

International patent applications WO2014/093936, WO2014/189805, WO2013/185052, U.S.2014/0341976, WO 2015/077354, WO2015/185565, PCT/IB2017/051945 and GB 1501462.4 disclose certain cyclic dinucleotides and their use in inducing immune responses via activation of STING. International patent application WO2017/106740 describes the use of cyclic dinucleotides and related scaffolds that measurably inhibit STING signaling and methods for identifying effective inhibitors of STING signaling. International patent applications WO 2017/175147 and WO 2017/175156 describe the use of heterocyclic amides and their analogs as STING modulators.

The compounds of the invention modulate the activity of STING and, therefore, may provide beneficial therapeutic effects in the treatment of diseases, disorders and/or conditions in which modulation of STING (stimulator of interferon genes) is beneficial, for example, for inflammatory, allergic and autoimmune diseases, metabolic diseases, cardiovascular diseases, infectious diseases, cancer, precancerous syndromes and as vaccine adjuvants.

Summary of the invention

The present invention relates to compounds according to formula (I):

in:

q is 0 or 1;

r is 0 or 1;

s is 0 or 1;

where q + r + s = 1 or 2;

When q is 0, ^RA1 and ^RA2 are each independently H, halogen, hydroxy, -OP(O)( ^OH ) ₂ , -OP(O)( ^{RIRII )} ₂ , -N( _Re )( ^Rf ), -CO2Rf, -N( ^{Rf ) CORb} , -N( ^Rg ) _SO2 ( _C1 - _C4alkyl )-N( ^Re )( ^Rf ),

-N(R ^g )CO(C ₁ -C ₄ alkyl)-N(R ^h )(R ^f ), optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino-, and optionally substituted (C ₁ -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-,

wherein the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino- and optionally substituted (C _{1 -C 6 alkyl)(C 1 -C 4 alkyl)amino-(C 1 -C 6 alkyl) are optionally substituted by 1-4 substituents each independently selected from the following substituents: hydroxy, -OP(O)(OH) 2 , -OP(O)(R I R II ) 2 , C 1 -C 4 alkoxy- , -N ( Re} ^{) (} _{R f )} ^{, -CO} ₂ (R f ), -CON(Re)(R ^f ), ^optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl or 5-6 membered heteroaryl are optionally substituted by 1-4 substituents each independently selected from the following substituents: C 1 -C 6 alkyl, optionally substituted (C 1 -C 6 alkyl)oxy-, optionally substituted ( ^C 1 -C 6 alkyl)amino-, and optionally substituted (C ₁ -C 6 alkyl) are optionally substituted by 1-4 substituents each independently selected from the following substituents: ₄ alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl)amino-, -(C ₁ -C ₆ alkyl)-NH ₂ , halo(C ₁ -C ₆ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C 1 -C _{4 alkoxy-, hydroxy-(C 2 -C 4 alkoxy ) -} , -(C _{2 -C 4} alkoxy)-OP(O)(R _I R ^{II )} ₂ R ^II ) ₂ , -C ₁ -C ₄ alkyl-(C ₁ -C ₄ alkoxy) and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

When r is 0, ^RB1 and ^RB2 are each independently H, optionally substituted _C1 - _C6 alkyl, halo( _C1 - _C6 alkyl), optionally substituted _C2 - _C6 alkenyl, optionally substituted C2- _C6 alkynyl, optionally substituted _{C3 - C6} cycloalkyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted phenyl, optionally substituted 5-6 membered heteroaryl or optionally substituted 9-10 membered heteroaryl,

wherein the optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₂ -C ₆ alkenyl, optionally substituted C ₂ -C ₆ alkynyl, optionally substituted C ₃ -C ₆ cycloalkyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted phenyl, optionally substituted 5-6 membered heteroaryl or optionally substituted 9-10 membered heteroaryl is optionally substituted with 1-4 substituents each independently selected from the group consisting of halogen, nitro, -R ^c , -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^c , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c , and -NR ^d SO ₂ R ^c ;

When s is 0, R ^C1 is H, halogen or C ₁ -C ₄ alkyl and R ^C2 is optionally substituted C ₁ -C ₄ alkyl, wherein the optionally substituted C ₁ -C ₄ alkyl is optionally substituted with a substituent selected from the group consisting of: -OR ^c , -NR ^c R ^d , -CO ₂ R ^c , -CONR ^c R ^d , -SO ₂ NR ^c R ^d and -OCONR ^c R ^d ;

When q is 1, ^RA1 and ^RA2 are each independently _-CH2- , ^-NRe- or -O-, and A together with ^RA1 and ^RA2 form a linking group, wherein A is -halo( _C1 - _C12alkyl )-, optionally substituted- _C1 - _C12alkyl- , optionally substituted- _C2 - _C12alkenyl- , optionally substituted- _C2 - _C12alkynyl- , optionally substituted- _C1 - _C6alkyl - _OC1 - _C6alkyl- , optionally substituted- _C1 - _C6alkyl -NRa ^- _C1 - _C6alkyl- , optionally substituted- _{C1 - C6alkyl-} ( _C3 - _C6cycloalkyl )-C1-C6alkyl-, optionally substituted- _{C1-C6alkyl-phenyl-C1 - C6alkyl- ,} optionally substituted- _C1 - _C6alkyl- (4-6memberedheterocycloalkyl)-C1- _{C6alkyl- 1} -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₆ alkyl-,

wherein the optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C The alkyl portion of the ₆ -alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C _{6 cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl portion of the optionally substituted -C 1 -C 6 alkyl-(C 3 -C 6 cycloalkyl)-C 1 -C 6 alkyl- , optionally} substituted -C ₁ -C 6 alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C _{1 -C 6 alkyl-(4-6-membered heterocycloalkyl)-C 1 -C 6 alkyl- or optionally substituted -C 1 -C 6 alkyl- ( 5-6 -} membered heteroaryl)-C ₁ -C ₆ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C 6 ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₁ -C ₄ alkoxy)-, -(C ₁ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

When r is 1, ^RB1 and ^RB2 are each independently ^{-CRdRf- ,} and B, together with ^RB1 and ^RB2, forms a linking group, wherein B is a bond or B is -halo( _C1 - _C10 alkyl)-, optionally substituted _-C1 - _C10 alkyl-, optionally substituted _-C2 - _C10 alkenyl-, optionally substituted _-C2 - _C10 alkynyl-, optionally substituted -C1 _{- C6} alkyl- _OC1 - _C6 alkyl-, optionally substituted _-C1 - _C6 alkyl- ^NRa - _C1 - _C6 alkyl-, optionally substituted _C3 - _C6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted _-C1 - _C4 alkyl-( _C3 - _C6 cycloalkyl) _-C1 - _C4 alkyl-, optionally substituted _-C1 -C6 -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl-, or optionally substituted -C ₁ -C ₄ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₄ alkyl-,

wherein the alkyl portion of the optionally substituted -C ₁ -C ₁₀ alkyl-, optionally substituted -C ₂ -C ₁₀ alkenyl-, optionally substituted -C ₂ -C ₁₀ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-(C ₃ -C ₆ cycloalkyl)-C 1 -C 4 alkyl-, optionally substituted -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C 4 alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl- or optionally substituted -C ₁ -C 4 alkyl-(5-6 membered heteroaryl-C 1 -C 4 alkyl)- is optionally substituted with 1 to 4 substituents each independently selected from the following substituents: -C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C 4 alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl- ₁ -C ₄ alkyl, halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ ,

-NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d ,

-SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c ,

-NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C ₆ cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl moiety of the optionally substituted C 3 -C 6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6-membered heterocycloalkyl, optionally substituted 5-6-membered heteroaryl, optionally substituted _{-C 1} -C ₄ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₄ alkyl- _, optionally substituted -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6-membered heterocycloalkyl)-C _{1 -C 4} alkyl- or optionally substituted -C ₁ -C ₄ alkyl-(5-6-membered heteroaryl)-C ₁ -C ₄ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the following: halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

When s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C together with R ^C1 and R ^C2 form a linking group, wherein C is -halo(C ₁ -C ₁₂ alkyl)-, optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C 6 ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₆ alkyl-,

wherein the optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C The alkyl portion of ₆ -alkyl- is optionally substituted by 1 or 2 substituents independently selected from the group consisting of halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ ,

-NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d ,

-SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c ,

-NR ^d SOR ^c , -NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C _{6 cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl portion of the optionally substituted -C 1 -C 6 alkyl-(C 3 -C 6 cycloalkyl)-C 1 -C 6 alkyl- , optionally} substituted -C ₁ -C 6 alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C _{1 -C 6 alkyl-(4-6-membered heterocycloalkyl)-C 1 -C 6 alkyl- or optionally substituted -C 1 -C 6 alkyl- ( 5-6 -} membered heteroaryl)-C ₁ -C ₆ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C 6 ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

R ³ and R ⁵ are each independently -CON(R ^d )(R ^f ), or one of R ³ and R ⁵ is -CON(R ^d )(R ^f ), and the other of R ³ and R ⁵ is H, COOH or -CO ₂ (R ^c );

^R4 and ^R6 are each independently selected from H, halogen, halo( _C1 - _C6 alkyl), halo( _C1 - _C6 alkoxy)-, hydroxy, -OP(O)(OH) ₂ , -OP( ^O )(R ^{I R II} ) ₂ , _-NH2 , -NRcRc, ^-NRcRd , ^-CORc , ^-CO2Rc , -N( ^{Rd ) CORc} , _-N (Rd)SO2Rc, -N( ^Rg ) _SO2 ( _C1 - _C2alkyl )-N( ^{Rh )} ( ^Rf ), -N ⁽ _Rg )CO( _C1 - _C2alkyl )-N( ^{Rh )} ( ^Rf ), optionally substituted ( _C1 - _C6alkyl ), optionally substituted ( _C1 - _C6alkyl )oxy-, optionally substituted ( _C1 -C6alkyl)amino-, and optionally substituted ( _C1 - _C6alkyl ) -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-,

wherein the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino- and optionally substituted (C ₁ -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-(C ₁ -C ₆ alkyl) are optionally substituted by 1 to 4 substituents each independently selected from the group consisting of -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^c , -NR ^c R ^d , -CO ₂ H , -CO ₂ R ^c , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH 2 ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c , -NR ^d SO ₂ R ^c , optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl or 5-6 membered heteroaryl is optionally substituted with 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d ;

R ¹⁴ is an optionally substituted C ₁ -C ₄ alkyl group, wherein the optionally substituted C ₁ -C ₄ alkyl group is optionally substituted by a substituent selected from the group consisting of: -OR ^c , -NR ^c R ^d , -CO ₂ R ^c , -CONR ^c R ^d , -SO ₂ NR ^c R ^d and -OCONR ^c R ^d ;

R ¹⁶ is H, halogen or C ₁ -C ₄ alkyl;

R ¹⁵ and R ¹⁷ are each independently H, cyclopropyl or C ₁ -C ₄ alkyl;

R ^a is H, -R ^c , -COR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ or -SO ₂ NR ^c R ^d ;

each R ^b is independently C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-OH, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , -(C ₁ -C ₄ alkyl)-O-(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-N(R ^e )(R ^f ), -(C ₁ -C ₄ alkyl)-O-CO(C ₁ -C ₄ alkyl), or -(C ₁ -C ₄ alkyl)-CO-O-(C ₁ -C ₄ alkyl);

each R ^c is independently C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-OH, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , -(C ₁ -C ₄ alkyl)-O-(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-N(R ^e )(R ^f ), -(C ₁ -C ₄ alkyl)-O-CO(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-CO-O-(C ₁ -C ₄ alkyl), optionally substituted C ₃ -C ₆ cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted 9-10 membered heteroaryl, optionally substituted -C ₁ -C ₄ alkyl-C ₃ -C ₆ cycloalkyl, optionally substituted -C ₁ -C ₄ alkyl-phenyl, optionally substituted -C ₁ -C ₄ alkyl-4-6 membered heterocycloalkyl, optionally substituted -C ₁ -C ₄ alkyl-5-6 membered heteroaryl or optionally substituted -C ₁ -C ₄ alkyl-9-10 membered heteroaryl,

wherein the substituted _C3 - _C6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted 9-10 membered heteroaryl, optionally substituted _-C1 - _C4 alkyl- _C3 - _C6 cycloalkyl, optionally substituted _-C1 - _C4 alkyl-phenyl, optionally substituted _-C1 - _C4 alkyl-4-6 membered heterocycloalkyl, optionally substituted _-C1 - _C4 alkyl-5-6 membered heteroaryl or optionally substituted -C1- _C4 alkyl-9-10 membered heteroaryl _{C3 -C6 cycloalkyl} , phenyl, 4-6 membered heterocycloalkyl, 5-6 membered heteroaryl or optionally substituted 9-10 membered heteroaryl moiety is optionally substituted with 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, -(C ₁ -C ₄ alkyl)NH ₂ , (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, -C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d ;

Each R ^d is independently H or C ₁ -C ₄ alkyl;

each R ^e is independently H, C ₁ -C ₄ alkyl, -CO(C ₁ -C ₄ alkyl), -OCO(C ₁ -C ₄ alkyl), -CO ₂ (C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)NH ₂ , -(C ₁ -C ₄ alkyl)C ₁ -C ₄ alkoxy, -CO-(optionally substituted 5-6 membered heterocycloalkyl), -CO(C ₁ -C ₄ alkyl)-(optionally substituted 5-6 membered heterocycloalkyl), -CO(optionally substituted 5-6 membered heteroaryl), or -CO(C ₁ -C ₄ alkyl)-(optionally substituted 5-6 membered heteroaryl),

wherein the optionally substituted 5-6 membered heterocycloalkyl or optionally substituted 5-6 membered heteroaryl is optionally substituted by 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-(C ₁ -C 4 ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d ;

Each R ^f is independently H or C ₁ -C ₄ alkyl;

^Rg and ^Rh are each independently H or _C1 - _C4 alkyl, or ^Rg and ^Rh together with the one or more atoms to which they are attached form a 5-6 membered ring;

and R ^I and R ^II are independently at each occurrence (C ₁ -C ₆ alkyl)oxy-; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof;

or a salt thereof.

It should be understood that references herein to compounds of formula (I) and their salts encompass compounds of formula (I) as free bases or salts thereof, e.g., as pharmaceutically acceptable salts thereof. Thus, in one embodiment, the present invention relates to compounds of formula (I) as free bases. In another embodiment, the present invention relates to compounds of formula (I) and their salts. In a further embodiment, the present invention relates to compounds of formula (I) and their pharmaceutically acceptable salts.

The compound according to formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt thereof, is a modulator of STING. Therefore, the present invention provides a compound of formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt thereof, for use in therapy. The present invention particularly provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as an active therapeutic substance in the treatment of a disease or condition mediated by STING, in particular for the treatment of a disease mediated by the agonist or antagonist of STING. The present invention also provides a compound of formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of a disease or condition mediated by STING.

The present invention also relates to a method for regulating STING, the method comprising contacting a cell with a compound according to formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt. The present invention further relates to a method for treating a disease or condition mediated by STING, the method comprising administering a therapeutically effective amount of a compound according to formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt, to a patient (human or other mammal, in particular human) in need thereof. Such STING-mediated diseases or conditions include inflammation, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, metabolic diseases and cardiovascular diseases. In addition, modulators of STING can be used as immunogenic compositions or vaccine adjuvants.

The present invention further relates to a pharmaceutical composition comprising a compound according to formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt, and a pharmaceutically acceptable excipient. Specifically, the present invention relates to a pharmaceutical composition for treating a STING-mediated disease or condition, wherein the composition comprises a compound according to formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt, and a pharmaceutically acceptable excipient.

DETAILED DESCRIPTION OF THE INVENTION

According to one aspect of the present invention, the present invention relates to a compound of formula (I)

in:

q is 0 or 1;

r is 0 or 1;

s is 0 or 1;

where q + r + s = 1 or 2;

When q is 0, ^RA1 and ^RA2 are each independently H, halogen, hydroxy, -OP(O)( ^OH ) ₂ , -OP(O)( ^{RIRII )} ₂ , -N( _Re )(Rf ⁾ , -CO2Rf, -N( ^{Rf ) CORb} , -N( ^Rg ) _SO2 ( _C1 - _C4 alkyl)-N( ^Re )( ^Rf ), -N( ^Rg )CO( _C1 - _C4 alkyl)-N( ^Rh )( ^Rf ), optionally substituted ( _C1 - _C6 alkyl), optionally substituted ( _C1 - _C6 alkyl)oxy-, optionally substituted ( _C1 - _C6 alkyl)amino-, and optionally substituted ( _C1 - _C6 alkyl)( _C1 - _C4 alkyl)amino-,

wherein the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino- and optionally substituted (C _{1 -C 6 alkyl)(C 1 -C 4 alkyl)amino-(C 1 -C 6 alkyl) are optionally substituted by 1-4 substituents each independently selected from the following substituents: hydroxy, -OP(O)(OH) 2 , -OP(O)(R I R II ) 2 , C 1 -C 4 alkoxy- , -N ( Re} ^{) (} _{R f )} ^{, -CO} ₂ (R f ), -CON(Re)(R ^f ), ^optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl or 5-6 membered heteroaryl are optionally substituted by 1-4 substituents each independently selected from the following substituents: C 1 -C 6 alkyl, optionally substituted (C 1 -C 6 alkyl)oxy-, optionally substituted ( ^C 1 -C 6 alkyl)amino-, and optionally substituted (C ₁ -C 6 alkyl) are optionally substituted by 1-4 substituents each independently selected from the following substituents: ₄ alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl)amino-, -(C ₁ -C ₆ alkyl)-NH ₂ , halo(C ₁ -C ₆ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C 1 -C _{4 alkoxy-, hydroxy-(C 2 -C 4 alkoxy ) -} , -(C _{2 -C 4} alkoxy)-OP(O)(R _I R ^{II )} ₂ R ^II ) ₂ , -C ₁ -C ₄ alkyl-(C ₁ -C ₄ alkoxy) and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

When r is 0, ^RB1 and ^RB2 are each independently H, optionally substituted _C1 - _C6 alkyl, halo( _C1 - _C6 alkyl), optionally substituted _C2 - _C6 alkenyl, optionally substituted C2- _C6 alkynyl, optionally substituted _{C3 - C6} cycloalkyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted phenyl, optionally substituted 5-6 membered heteroaryl or optionally substituted 9-10 membered heteroaryl,

wherein the optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₂ -C ₆ alkenyl, optionally substituted C ₂ -C ₆ alkynyl, optionally substituted C ₃ -C ₆ cycloalkyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted phenyl, optionally substituted 5-6 membered heteroaryl or optionally substituted 9-10 membered heteroaryl is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, nitro, -R ^c , -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^c , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c , and -NR ^d SO ₂ R ^c ;

When s is 0, R ^C1 is H, halogen or C ₁ -C ₄ alkyl, and R ^C2 is optionally substituted C ₁ -C ₄ alkyl, wherein the optionally substituted C ₁ -C ₄ alkyl is optionally substituted with a substituent selected from the group consisting of: -OR ^c , -NR ^c R ^d , -CO ₂ R ^c , -CONR ^c R ^d , -SO ₂ NR ^c R ^d and -OCONR ^c R ^d ;

When q is 1, ^RA1 and ^RA2 are each independently _-CH2- , ^-NRe- or -O-, and A together with ^RA1 and ^RA2 form a linking group, wherein A is -halo( _C1 - _C12alkyl )-, optionally substituted- _C1 - _C12alkyl- , optionally substituted- _C2 - _C12alkenyl- , optionally substituted- _C2 - _C12alkynyl- , optionally substituted- _C1 - _C6alkyl - _OC1 - _C6alkyl- , optionally substituted- _C1 - _C6alkyl -NRa ^- _C1 - _C6alkyl- , optionally substituted- _{C1 - C6alkyl-} ( _C3 - _C6cycloalkyl )-C1-C6alkyl-, optionally substituted- _{C1-C6alkyl-phenyl-C1 - C6alkyl- ,} optionally substituted- _C1 - _C6alkyl- (4-6memberedheterocycloalkyl)-C1- _{C6alkyl- 1} -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₆ alkyl-,

wherein the optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C The alkyl portion of the ₆ -alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C _{6 cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl portion of the optionally substituted -C 1 -C 6 alkyl-(C 3 -C 6 cycloalkyl)-C 1 -C 6 alkyl- , optionally} substituted -C ₁ -C 6 alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C _{1 -C 6 alkyl-(4-6-membered heterocycloalkyl)-C 1 -C 6 alkyl- or optionally substituted -C 1 -C 6 alkyl- ( 5-6 -} membered heteroaryl)-C ₁ -C ₆ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C 6 ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₁ -C ₄ alkoxy)-, -(C ₁ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

When r is 1, ^RB1 and ^RB2 are each independently ^{-CRdRf- ,} and B, together with ^RB1 and ^RB2, forms a linking group, wherein B is a bond or B is -halo( _C1 - _C10 alkyl)-, optionally substituted _-C1 - _C10 alkyl-, optionally substituted _-C2 - _C10 alkenyl-, optionally substituted _-C2 - _C10 alkynyl-, optionally substituted -C1 _{- C6} alkyl- _OC1 - _C6 alkyl-, optionally substituted _-C1 - _C6 alkyl- ^NRa - _C1 - _C6 alkyl-, optionally substituted _C3 - _C6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted _-C1 - _C4 alkyl-( _C3 - _C6 cycloalkyl) _-C1 - _C4 alkyl-, optionally substituted _-C1 -C6 -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl-, or optionally substituted -C ₁ -C ₄ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₄ alkyl-,

wherein the alkyl portion of the optionally substituted -C ₁ -C ₁₀ alkyl-, optionally substituted -C ₂ -C ₁₀ alkenyl-, optionally substituted -C ₂ -C ₁₀ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₄ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl- or optionally substituted -C ₁ -C ₄ alkyl-(5-6 membered heteroaryl-C ₁ -C ₄ alkyl)- is optionally substituted with 1-C 4 alkyl- wherein the at least one aryl group is substituted with 4 substituents each independently selected from the group consisting of -C ₁ -C ₄ alkyl, halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C ₆ cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl moiety of the optionally substituted C 3 -C 6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6-membered heterocycloalkyl, optionally substituted 5-6-membered heteroaryl, optionally substituted _{-C 1} -C ₄ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₄ alkyl- _, optionally substituted -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6-membered heterocycloalkyl)-C _{1 -C 4} alkyl- or optionally substituted -C ₁ -C ₄ alkyl-(5-6-membered heteroaryl)-C ₁ -C ₄ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the following: halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

When s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C together with R ^C1 and R ^C2 form a linking group, wherein C is -halo(C ₁ -C ₁₂ alkyl)-, optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C 6 ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₆ alkyl-,

wherein the optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C The alkyl portion of the ₆ -alkyl- is optionally substituted with 1 or 2 substituents each independently selected from the group consisting of halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c , and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C _{6 cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl portion of the optionally substituted -C 1 -C 6 alkyl-(C 3 -C 6 cycloalkyl)-C 1 -C 6 alkyl- , optionally} substituted -C ₁ -C 6 alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C _{1 -C 6 alkyl-(4-6-membered heterocycloalkyl)-C 1 -C 6 alkyl- or optionally substituted -C 1 -C 6 alkyl- ( 5-6 -} membered heteroaryl)-C ₁ -C ₆ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C 6 ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

R ³ and R ⁵ are each independently -CON(R ^d )(R ^f ), or one of R ³ and R ⁵ is -CON(R ^d )(R ^f ), and the other of R ³ and R ⁵ is H, COOH or -CO ₂ (R ^c );

^R4 and ^R6 are each independently selected from H, halogen, halo( _C1 - _C6 alkyl), halo( _C1 - _C6 alkoxy)-, hydroxy, -OP(O)(OH) ₂ , -OP( ^O )(R ^{I R II} ) ₂ , _-NH2 , -NRcRc, ^-NRcRd , ^-CORc , ^-CO2Rc , -N( ^{Rd ) CORc} , _-N (Rd)SO2Rc, -N( ^Rg ) _SO2 ( _C1 - _C2alkyl )-N( ^{Rh )} ( ^Rf ), -N ⁽ _Rg )CO( _C1 - _C2alkyl )-N( ^{Rh )} ( ^Rf ), optionally substituted ( _C1 - _C6alkyl ), optionally substituted ( _C1 - _C6alkyl )oxy-, optionally substituted ( _C1 -C6alkyl)amino-, and optionally substituted ( _C1 - _C6alkyl ) -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-,

wherein the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino- and optionally substituted (C ₁ -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-(C ₁ -C ₆ alkyl) are optionally substituted by 1 to 4 substituents each independently selected from the group consisting of -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^c , -NR ^c R ^d , -CO ₂ H , -CO ₂ R ^c , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH 2 ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c , -NR ^d SO ₂ R ^c , optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl or 5-6 membered heteroaryl is optionally substituted with 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , -C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d ;

R ¹⁴ is an optionally substituted C ₁ -C ₄ alkyl group, wherein the optionally substituted C ₁ -C ₄ alkyl group is optionally substituted by a substituent selected from the group consisting of: -OR ^c , -NR ^c R ^d , -CO ₂ R ^c , -CONR ^c R ^d , -SO ₂ NR ^c R ^d and -OCONR ^c R ^d ;

R ¹⁶ is H, halogen or C ₁ -C ₄ alkyl;

R ¹⁵ and R ¹⁷ are each independently H, cyclopropyl or C ₁ -C ₄ alkyl;

R ^a is H, -R ^c , -COR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ or -SO ₂ NR ^c R ^d ;

each R ^b is independently C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-OH, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , -(C ₁ -C ₄ alkyl)-O-(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-N(R ^e )(R ^f ), -(C ₁ -C ₄ alkyl)-O-CO(C ₁ -C ₄ alkyl), or -(C ₁ -C ₄ alkyl)-CO-O-(C ₁ -C ₄ alkyl);

each R ^c is independently C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-OH, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , -(C ₁ -C ₄ alkyl)-O-(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-N(R ^e )(R ^f ), -(C ₁ -C ₄ alkyl)-O-CO(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-CO-O-(C ₁ -C ₄ alkyl), optionally substituted C ₃ -C ₆ cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted 9-10 membered heteroaryl, optionally substituted -C ₁ -C ₄ alkyl-C ₃ -C ₆ cycloalkyl, optionally substituted -C ₁ -C ₄ alkyl-phenyl, optionally substituted -C ₁ -C ₄ alkyl-4-6 membered heterocycloalkyl, optionally substituted -C ₁ -C ₄ alkyl-5-6 membered heteroaryl or optionally substituted -C ₁ -C ₄ alkyl-9-10 membered heteroaryl,

wherein the substituted _C3 - _C6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted 9-10 membered heteroaryl, optionally substituted _-C1 - _C4 alkyl- _C3 - _C6 cycloalkyl, optionally substituted _-C1 - _C4 alkyl-phenyl, optionally substituted _-C1 - _C4 alkyl-4-6 membered heterocycloalkyl, optionally substituted _-C1 - _C4 alkyl-5-6 membered heteroaryl or optionally substituted -C1- _C4 alkyl-9-10 membered heteroaryl _{C3 -C6 cycloalkyl} , phenyl, 4-6 membered heterocycloalkyl, 5-6 membered heteroaryl or optionally substituted 9-10 membered heteroaryl moiety is optionally substituted with 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, -(C ₁ -C ₄ alkyl)NH ₂ , (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, -C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d ;

Each R ^d is independently H or C ₁ -C ₄ alkyl;

each R ^e is independently H, C ₁ -C ₄ alkyl, -CO(C ₁ -C ₄ alkyl), -OCO(C ₁ -C ₄ alkyl), -CO ₂ (C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)NH ₂ , -(C ₁ -C ₄ alkyl)C ₁ -C ₄ alkoxy, -CO-(optionally substituted 5-6 membered heterocycloalkyl), -CO(C ₁ -C ₄ alkyl)-(optionally substituted 5-6 membered heterocycloalkyl), -CO(optionally substituted 5-6 membered heteroaryl), or -CO(C ₁ -C ₄ alkyl)-(optionally substituted 5-6 membered heteroaryl),

wherein the optionally substituted 5-6 membered heterocycloalkyl or optionally substituted 5-6 membered heteroaryl is optionally substituted by 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-(C ₁ -C 4 ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d ;

Each R ^f is independently H or C ₁ -C ₄ alkyl;

^Rg and ^Rh are each independently H or _C1 - _C4 alkyl, or ^Rg and ^Rh together with the one or more atoms to which they are attached form a 5-6 membered ring;

and R ^I and R ^II are independently at each occurrence (C ₁ -C ₆ alkyl)oxy-; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof;

or a salt thereof.

The alternative definitions of the various groups and substituents of formula (I) provided throughout the specification are intended to specifically describe each compound species disclosed herein individually, as well as groups of one or more compound species. The scope of the present invention includes any combination of these group and substituent definitions. The compounds of the present invention are only those considered to be "chemically stable" as understood by those skilled in the art.

It will be appreciated by those skilled in the art that the compounds of the present invention may exist in other tautomeric forms (including zwitterionic forms) or isomeric forms. All tautomeric (including zwitterionic forms) and isomeric forms of the chemical formulae and compounds described herein are intended to be encompassed within the scope of the present invention.

Those skilled in the art will also appreciate that the compounds of the present invention may exist in tautomeric (or isomeric) forms (including but not limited to Formula (A), Formula (B) and/or Formula (C)) or zwitterionic forms (including but not limited to Formula (D) or Formula (E)). In Formula (B), (C), (D) or (E), each occurrence of R is independently H or any suitable substituent on nitrogen, such as an alkyl group.

The chemical names provided for the intermediate compounds and/or compounds of the invention described herein may refer to any tautomeric/isomeric representation of such compounds (in some cases, such alternative names are provided by the experiments). It should be understood that any reference to a named compound (an intermediate compound or compound of the invention) or a structurally depicted compound (an intermediate compound or compound of the invention) is intended to encompass all tautomeric/isomeric forms of such compounds, including zwitterionic forms, and any mixtures thereof.

As used herein, the term "alkyl" represents a saturated, straight or branched hydrocarbon group having a specified number of carbon atoms. The term " _C1 - _C4 alkyl" refers to a straight or branched alkyl moiety containing 1 to 4 carbon atoms. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

When a substituent term such as "alkyl" is used in combination with another substituent term, for example as in "hydroxy(C ₁ -C ₄ alkyl)", the linking substituent term (e.g., alkyl) is intended to encompass a divalent moiety where the point of attachment is through that linking substituent. Examples of "hydroxy(C ₁ -C ₄ alkyl)" groups include, but are not limited to, hydroxymethyl, hydroxyethyl, and hydroxyisopropyl.

As used herein, the term "halo(alkyl)" represents a saturated, straight-chain or branched hydrocarbon radical having a specified number (n) of carbon atoms and one or more (up to 2n + 1) halogen atoms. For example, the term "halo(C ₁ -C ₄ )alkyl" represents a radical having one or more halogen atoms, which may be the same or different, at one or more carbon atoms of an alkyl moiety containing 1 to 4 carbon atoms. Examples of "halo(C ₁ -C ₄ alkyl)" radicals include, but are not limited to, -CF ₃ (trifluoromethyl), -CCl ₃ (trichloromethyl), 1,1-difluoroethyl, 2,2,2-trifluoroethyl and hexafluoroisopropyl.

"Alkenyl" refers to a straight or branched chain hydrocarbon group having the specified number of carbon atoms and at least 1 and up to 3 carbon-carbon double bonds. Examples include ethenyl and propenyl.

"Alkynyl" refers to a straight or branched chain hydrocarbon group having the specified number of carbon atoms and at least 1 and up to 3 carbon-carbon triple bonds. Examples include ethynyl and propynyl.

"Alkoxy-" or "(alkyl)oxy-" refers to an "alkyl-oxy-" group containing an alkyl moiety having the specified number of carbon atoms bonded through an oxygen linking atom. For example, the term "C ₁ -C ₄ alkoxy-" represents a saturated, straight-chain or branched hydrocarbon moiety having at least 1 and up to 4 carbon atoms bonded through an oxygen linking atom. Exemplary "C ₁ -C ₄ alkoxy-" or "(C ₁ -C ₄ alkyl)oxy-" groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, and tert-butoxy.

As used herein, the term "halo(alkoxy)-" represents a saturated, straight-chain or branched hydrocarbon radical having a specified number (n) of carbon atoms and one or more (up to 2n+1) halogen atoms bonded through an oxygen linking atom. For example, the term "halo(C ₁ -C ₄ alkoxy)-" refers to a "haloalkyl-oxy-" radical containing a "halo(C ₁ -C ₄ alkyl)" moiety bonded through an oxygen linking atom. Exemplary "halo(C ₁ -C ₄ alkoxy)-" radicals include, but are not limited to, -OCHF ₂ (difluoromethoxy), -OCF ₃ (trifluoromethoxy), -OCH ₂ CF ₃ (trifluoroethoxy), and -OCH(CF ₃ ) ₂ (hexafluoroisopropoxy).

A carbocyclic group or moiety is a cyclic group or moiety in which the ring members are carbon atoms, which may be saturated, partially unsaturated (non-aromatic) or fully unsaturated (aromatic).

"Cycloalkyl" refers to a non-aromatic, saturated, hydrocarbon ring group containing the specified number of carbon atoms in the ring. For example, the term "C ₃ -C ₆ cycloalkyl" refers to a cyclic group having three to six ring carbon atoms. Exemplary "C ₃ -C ₆ cycloalkyl" groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

A heterocyclic group or moiety is a cyclic group or moiety having atoms of at least two different elements as ring members, which cyclic group or moiety may be saturated, partially unsaturated (non-aromatic) or fully unsaturated (aromatic).

"Heteroatom" refers to a nitrogen, sulfur or oxygen atom, for example a nitrogen atom or an oxygen atom.

"Heterocycloalkyl" refers to a non-aromatic, monocyclic or bicyclic group containing 3-10 ring atoms and containing one or more (usually one or two) heteroatom ring members independently selected from oxygen, sulfur and nitrogen. The point of attachment of the heterocycloalkyl group can be any suitable carbon or nitrogen atom.

Examples of “heterocycloalkyl” groups include, but are not limited to, aziridine, thiirane, oxirane, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, 1,3-dioxanyl, 1,4-dioxanyl, 1,3-oxathiolanyl, 1,3-dithiolanyl, 1,4-oxathiolanyl, 1,4-dithiolanyl, morpholinyl, thiomorpholinyl, and hexahydro- 1H -1,4-diazepinyl.

Examples of "4-membered heterocycloalkyl" groups include oxetanyl, thietanyl and azetidinyl.

The term "5-6 membered heterocycloalkyl" represents a saturated, monocyclic group containing 5 or 6 ring atoms, including one or two heteroatoms independently selected from oxygen, sulfur and nitrogen. Illustrative examples of 5 to 6 membered heterocycloalkyl groups include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, piperazinyl, morpholinyl and thiomorpholinyl.

"Heteroaryl" refers to an aromatic monocyclic or bicyclic group containing 5 to 10 ring atoms, including 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein at least a portion of the group is aromatic. For example, the term encompasses a bicyclic heterocyclic-aryl group containing a phenyl ring fused to a heterocyclic portion or a heteroaryl ring portion fused to a carbocyclic portion. The point of attachment of the heteroaryl group may be any suitable carbon or nitrogen atom.

The term "5-6 yuan heteroaryl" represents an aromatic monocyclic group containing 5 or 6 ring atoms, which includes at least one carbon atom and 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur. The selected 5-yuan heteroaryl group contains a nitrogen, oxygen or sulfur ring heteroatom, and optionally contains 1,2 or 3 additional nitrogen ring atoms. The selected 6-yuan heteroaryl group contains 1,2 or 3 nitrogen ring heteroatoms. The example of the 5-yuan heteroaryl group includes furyl (furyl) (furanyl (furanyl)), thienyl, pyrrolyl, imidazoles, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl and oxadiazolyl. The selected 6-yuan heteroaryl group includes pyridyl (pyridinyl) (pyridyl (pyridyl)), pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl.

The term "9-10 membered heteroaryl" refers to an aromatic bicyclic group containing 9 or 10 ring atoms, including 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur. Examples of 9-membered heteroaryl (6,5-fused heteroaryl) include benzothiophenyl, benzofuranyl, indolyl, indolinyl (dihydroindolinyl), isoindolyl, isoindolyl, indazolyl, isobenzofuranyl, 2,3-dihydrobenzofuranyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzimidazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, imidazopyridyl, pyrazolopyridyl, triazolopyridyl and 1,3-benzodioxolyl.

Examples of 10-membered heteroaryl (6,6-fused heteroaryl) groups include quinolinyl (quinolyl), isoquinolinyl, phthalazinyl, naphthyridinyl (1,5-naphthyridinyl, 1,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl), quinazolinyl, quinoxalinyl, 4H-quinolizinyl, 1,2,3,4-tetrahydroquinolinyl (tetrahydroquinolinyl), 1,2,3,4-tetrahydroisoquinolinyl (tetrahydroisoquinolinyl), cinnolinyl, pteridinyl, and 2,3-dihydrobenzo[b][1,4]dioxinyl.

The terms "halogen" and "halo" refer to a halogen radical, such as a fluorine, chlorine, bromine or iodine substituent.

"Oxo" represents the oxygen portion of a double bond; for example, if directly bound to a carbon atom it would form a carbonyl group (C=O).

"Hydroxy" or "hydroxyl" is intended to mean the group -OH.

As used herein, the term "cyano" refers to a nitrile group, -C≡N.

As used herein, the term "optionally substituted" means that a group (such as an alkyl, cycloalkyl, alkoxy, heterocycloalkyl, aryl or heteroaryl group) or a ring or a moiety may not be substituted, or the group, ring or moiety may be substituted with one or more substituents as defined in the substituent definitions (A, ^R3, etc.) provided herein. In the case where a group can be selected from a number of alternative groups, the selected groups may be the same or different.

The term "independently" means that when more than one substituent is selected from a number of possible substituents, those substituents may be the same or different.

The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

As used herein, the term "compound(s) of the invention" or "compound(s) of this invention" means a compound of formula (I) as defined herein, in any form, i.e. any tautomeric/isomerized form, any salt or non-salt form (e.g. as a free acid or base form, or as a salt, in particular a pharmaceutically acceptable salt thereof), and in any physical form thereof (e.g. including non-solid forms (e.g. liquid or semi-solid forms), as well as solid forms (e.g. amorphous or crystalline forms, specific polymorphic forms, solvate forms (including hydrate forms (e.g. monohydrates, dihydrates and hemihydrates))), as well as mixtures of various forms.

Therefore, included in the present invention are compounds of formula (I) as defined herein, in any salt or non-salt form and any physical form thereof, as well as mixtures of various forms. Although so included in the present invention, it is understood that compounds of formula (I) as defined herein, in any salt or non-salt form thereof and in any physical form, may have different activity levels, different bioavailabilities and different handling characteristics for formulation purposes.

In one embodiment of the compounds of the invention, ^R3 and ^R5 are each independently -CON( ^Rd )( ^Rf ), or one of ^R3 and ^R5 is -CON( ^Rd )( ^Rf ), and the other of ^R3 and ^R5 is H or _-CO2 ( ^Rc ). In one embodiment, ^R3 and ^R5 are each independently -CON( ^Rd )( ^Rf ). In another embodiment, one of ^R3 and ^R5 is -CON( ^Rd )( ^Rf ) and the other of ^R3 and ^R5 is H. In a specific embodiment, ^R3 and ^R5 are each _-CONH2 .

It is understood that when q is 0, A does not exist and ^RA1 and ^RA2 are not connected. Similarly, it is understood that when r is 0, B does not exist and ^RB1 and ^RB2 are not connected. Similarly, it is understood that when s is 0, C does not exist and ^RC1 and ^RC2 are not connected.

In one embodiment of the compound of the present invention, q is 1, r is 0 and s is 0 (q+r+s=1), and the compound has formula (I-A) or (I-a):

。

.

In one embodiment of the compounds of the present invention, q is 0, r is 1 and s is 0 (q+r+s=1), and the compound has formula (I-B) or (I-b):

。

.

In one embodiment of the compound of the present invention, q is 0, r is 0 and s is 1 (q+r+s=1), and the compound has formula (I-C) or (I-c):

。

.

In one embodiment of the compounds of the present invention, q is 1, r is 1 and s is 0 (q+r+s=2), and the compound has the formula (I-AB) or (I-ab):

。

.

In one embodiment of the compounds of the present invention, q is 1, r is 0 and s is 1 (q+r+s=2), and the compound has the formula (I-AC) or (I-ac):

。

.

In one embodiment of the compounds of the present invention, q is 0, r is 1 and s is 1 (q+r+s=2), and the compound has the formula (I-BC) or (I-bc):

。

.

In one embodiment of the compounds of the present invention, q is 0, and ^RA1 and ^RA2 are each independently H, halogen, hydroxy, -OP( _O )(OH) ₂ , -OP ⁽ O)( ^RIRII ) ₂ , -N( ^{Re )} ( ^Rf ) ^, -CO2Rf, -N( ^Rf )CORb, -N( ^Rg ) _SO2 ( ^C1 -C4alkyl)-N(Re)( ^Rf ), -N ₍ ^Rg )CO( _{C1 - C4alkyl} )-N( ^Rh )( ^Rf ), optionally substituted ( _C1 - _C6alkyl ), optionally substituted ( _C1 - _C6alkyl )oxy-, optionally substituted ( _C1 - _C6alkyl )amino-, and optionally substituted ( _C1 - _C6alkyl )( _C1 - _C4alkyl )amino-,

wherein the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino- and optionally substituted (C ₁ -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-(C ₁ -C ₆ alkyl) are optionally substituted by 1 to 4 substituents each independently selected from the group consisting of hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-, -N( ^Re )(R ^f ), -CO ₂ (R ^f ), -CON( ^Re )(R ^f ), optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl or 5-6 membered heteroaryl is optionally substituted by 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl)amino-, -(C ₁ -C ₆ alkyl)-NH2, halo(C ₁ -C ₆ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , _- (C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, _C 1 -C 4 alkoxy -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-.

In one embodiment of the compounds of the present invention, q is 0 and ^RA1 and ^RA2 are each independently H, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl), hydroxy(C ₁ -C ₄ alkyl)-, amino(C ₁ -C ₄ alkyl)-, (C ₁ -C ₄ alkyl)amino(C ₁ -C ₄ alkyl)-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino(C ₁ -C ₄ alkyl)-, C ₁ -C ₄ alkoxy-, hydroxy(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R I R ^II) R ^II ) ₂ , amino(C ₂ -C ₄ alkoxy)-, (C ₁ -C ₄ alkyl)amino(C ₂ -C ₄ alkoxy)-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino(C ₂ -C ₄ alkoxy)-, 6-membered heterocycloalkyl-(C ₁ -C ₄ alkyl)-, phenyl(C ₁ -C ₄ alkoxy)-, (C ₁ -C ₄ alkyl)OCONH(C ₁ -C ₄ alkyl)-, hydroxy(C ₁ -C ₄ alkyl)amino-, -amino(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -amino(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , (C ₁ -C ₄ alkyl)CONH-, (C ₁ -C ₄ alkyl)CON(C ₁ -C ₄ alkyl)-, -CO ₂ H, -CO ₂ (C ₁ -C ₄ alkyl), amino (C ₁ -C ₄ alkyl) CONH-, (C ₁ -C ₄ alkyl) amino (C ₁ -C ₄ alkyl) CONH-, amino (C ₁ -C ₄ alkyl) CON (C ₁ -C ₄ alkyl) -, (C ₁ -C ₄ alkyl) amino (C ₁ -C ₄ alkyl) CONH-, amino (C ₁ -C ₄ alkyl) CON (C ₁ -C ₄ alkyl) -, (C ₁ -C ₄ alkyl) amino (C ₁ -C ₄ alkyl) CON (C 1 -C 4 alkyl) -, hydroxy (C ₁ -C ₄ alkyl) CONH-, -NHCO (C ₁ -C ₄ alkyl) -OP (O) (OH) ₂ , -NHCO (C ₁ -C ₄ alkyl) -OP (O) (R ^I R ^II ) ₂ , (C ₁ -C ₄ alkyl) (C ₁ -C ₄ alkyl) amino (C ₁ -C ₄ alkyl) CON (C ₁ -C ₄ alkyl)-, hydroxy(C ₁ -C ₄ alkyl)CON(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)NCO(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)NCO(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , HO ₂ C(C ₁ -C ₄ alkoxy)-, (C ₁ -C ₄ alkyl)OCO(C ₁ -C ₄ alkoxy)-, H ₂ NCO(C ₁ -C ₄ alkoxy)-, (C ₁ -C ₄ alkyl)HNCO(C ₁ -C ₄ alkoxy)-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)NCO(C ₁ -C ₄ alkoxy)-, and -NHSO ₂ (C ₁ -C ₄ alkyl).

In one embodiment, q is 0 and ^RA1 and ^RA2 are each independently H, (C ₁ -C ₆ alkyl)oxy- or hydroxy(C ₂ -C ₆ alkyl)oxy-. In one embodiment, q is 0 and ^RA1 and ^RA2 are each independently H, (C ₁ -C ₆ alkyl)oxy-, hydroxy(C ₂ -C ₆ alkyl)oxy-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ . In one embodiment, q is 0 and ^RA1 and ^RA2 are each H. In selected embodiments, q is 0 and ^RA1 and ^RA2 are independently selected from H, -OCH ₂ CH ₂ CH ₂ OH and -OCH ₃ .

In one embodiment, q is 0 and ^RA2 and ^RA1 are each independently H, optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy-, wherein the C1- _C6 alkyl of the optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _{C1 - C6} alkyl)oxy- is optionally substituted with 1-4 substituents each independently selected from the group consisting of hydroxy, -OP(O)(OH) ₂ , -OP(O)( ^{R1RII )} ₂ , _C1 - _C4 alkoxy, -N( ^Re )( ^Rf ), -COOH, optionally substituted phenyl and optionally substituted 5-6 membered heterocycloalkyl, and each ^Re is independently selected from the group consisting of H, _C1 - _C4 alkyl, -CO( _C1 - _C4 alkyl), -OCO( _C1 - _C4 alkyl), -( _C1 - _C4 alkyl) _NH2 , -( _C1 - _C4 alkyl) C ₁ -C ₄ alkoxy or -CO ₂ (C ₁ -C ₄ alkyl).

In one embodiment, q is 0 and R ^A2 and R ^A1 are each independently H, optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-, and the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy

The C ₁ -C ₆ alkyl group is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -N( ^Re )( ^Rf ), C ₁ -C ₄ alkoxy, phenyl, and an optionally substituted 5-6 membered heterocycloalkyl group containing at least one nitrogen or oxygen as a ring member, and each ^{Re is} each independently selected from the group consisting of H, C ₁ -C ₄ alkyl, -(C ₁ -C ₄ alkyl)NH ₂ or -(C ₁ -C ₄ alkyl)C ₁ -C ₄ alkoxy.

In one embodiment, q is 0 and at least one of ^RA2 or ^RA1 is each independently H, optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1-C6 alkyl of the optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy- is optionally substituted with 1-4 substituents each independently selected from the group consisting of -N ₍ ^Re )( ^Rf ), tetrahydropyran, pyrrolidinyl, piperazinyl, piperidinyl and morpholinyl, and each ^{Re is} each independently selected from H, _C1 - _C4 alkyl, -( _C1 - _C4 alkyl) _NH2 or -( _{C1 - C4} alkyl) _C1 - _C4 alkoxy.

In one embodiment, q is 0 and at least one of ^RA2 or ^RA1 is each independently H, optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1- _C6 alkyl of the optionally substituted ( _C1 -C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy- is optionally substituted with 1-4 substituents each independently selected from the following substituents: tetrahydropyran, pyrrolidinyl, piperazinyl, piperidinyl and morpholinyl, and each ^Re is each independently selected from H or _{C1 - C4 alkyl} .

In one embodiment, q is 0 and ^RA2 and ^RA1 are each independently selected from H, hydroxy or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the _C1 - _C6 alkyl of the optionally substituted ( _C1 - _C6 alkyl)oxy- is optionally substituted with hydroxy, phenyl or morpholinyl, wherein phenyl or morpholinyl are each independently substituted with methyl or methoxy.

In one embodiment, q is 0, ^RA1 and ^RA2 are independently H or optionally substituted ( _C1 - _C6 alkyl)oxy-, wherein the alkyl of said optionally substituted ( _C1 - _C6 alkyl)oxy- is optionally substituted with one substituent selected from the group consisting of hydroxy, optionally substituted phenyl and optionally substituted 5-6 membered heterocycloalkyl, wherein said phenyl and 5-6 membered heterocycloalkyl are optionally substituted with one substituent selected from the group consisting of _{C1- C3} alkyl and _C1 - _C3 alkoxy.

In one embodiment, q is 0, ^RA1 and ^RA2 are independently H or optionally substituted ( _C1 - _C6 alkyl)oxy-, wherein the alkyl of the optionally substituted ( _C1 - _C6 alkyl)oxy- is optionally substituted with one substituent selected from the group consisting of hydroxy, optionally substituted phenyl and optionally substituted morpholinyl, wherein the phenyl and morpholinyl are optionally substituted with one substituent selected from the group consisting of _C1 - _C3 alkyl and _C1 - _C3 alkoxy.

In one embodiment, q is 0, R ^A1 and R ^A2 are independently H, hydroxy or optionally substituted (C ₁ -C ₁₂ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₁₂ alkyl)oxy-group is optionally substituted by 1 to 4 substituents independently selected from the following: hydroxyl, COOH and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1 to 4 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, q is 0, one of ^RA1 and ^RA2 is H and the other of ^RA1 and ^RA2 is hydroxy or optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted by 1-2 substituents independently selected from the following: hydroxyl and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted with 1 to 2 substituents each independently being (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, q is 0 and both ^RA1 and ^RA2 are H.

In one embodiment, r is 0 and ^RB1 and ^RB2 are each H.

In another embodiment, r is 0 and ^RB1 and ^RB2 are each independently H, optionally substituted _C1 - _C6 alkyl, halo( _C1 - _C6 alkyl), optionally substituted _C2 - _C6 alkenyl, optionally substituted _C2 - _C6 alkynyl, optionally substituted _C3 - _C6 cycloalkyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, or optionally substituted 9 membered heteroaryl.

In one embodiment of the compounds of the invention, s is 0, and R ^C1 is H, halogen or C ₁ -C ₄ alkyl, and R ^C2 is optionally substituted C ₁ -C ₄ alkyl, wherein said optionally substituted C ₁ -C ₄ alkyl is optionally substituted with a substituent selected from the group consisting of -OR ^c , -NR ^c R ^d , -CO ₂ R ^c , -CONR ^c R ^d , -SO ₂ NR ^c R ^d and -OCONR ^c R ^d .

In one embodiment of the compounds of the present invention, when s is 0, R ^C1 is H and R ^C2 is C ₁ -C ₄ alkyl. In another embodiment, when s is 0, R ^C1 is C ₁ -C ₃ alkyl, especially methyl. In another embodiment, when s is 0, R ^C2 is C ₁ -C ₃ alkyl, especially methyl or ethyl. In a selected embodiment, when s is 0, R ^C2 is ethyl.

In one embodiment, q is 1 and A, together with ^RA1 and ^RA2, forms a 4-8 membered linking group. In another embodiment, q is 1 and A, together with ^RA1 and ^RA2, forms a 4-6 membered linking group. In yet another embodiment, q is 1 and A, together with ^RA1 and ^RA2, forms a 5-membered linking group.

In another embodiment, q is 1, ^RA1 and ^RA2 are each independently _-CH2- , ^-NRe- or -O-, and A is a substituted _-C2 - _C10 alkyl- group or an unsubstituted _-C2 - _C10 alkyl-, -C2-C10 alkenyl-, -C2- _C10 alkynyl-, -C1 _{- C4} alkyl-OC1 _{- C4} alkyl- or _-C1 - _C4 alkyl-NRa ^- _C1 - _C4 alkyl- group, the substituted _-C2 - _C10 alkyl- group being substituted with 1-4 substituents each independently selected from _the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP ₍ O)(R ^I R ^II ) ₂ , amino, (C1- _C4 alkyl)amino-, ( _C1 - _C4 alkyl)( _{C1 -C4 alkyl} )amino- _{, halo(C1 -} C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, q is 1, ^RA1 and ^RA2 are each independently _-CH2- , ^-NRe- or -O-, and A is a substituted _-C2 - _C8 alkyl-group or an unsubstituted -C2- _C8 alkyl-, -C2-C8 alkenyl-, -C2- _C8 alkynyl-, _-C1 - _{C2 alkyl} -OC1 _{- C2} alkyl- or -C1 _{-C2 alkyl} -NRa ^- _{C1 - C2} alkyl-group, the substituted _{-C2- C8} alkyl-group being substituted with 1-2 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH _{) 2} , -OP(O)(R ^I R ^II ) ₂ , amino, ( _C1 - _C4 alkyl)amino-, ( _C1 - _C4 alkyl)( _C1 - _{C4 alkyl} )amino-, halo( _C1 -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, q is 1, ^RA1 and ^RA2 are each independently _-CH2- , ^-NRe- or -O-, and A is a substituted _-C2 - _C6 alkyl- group or an unsubstituted _-C2 - _C6 alkyl-, -C2-C6 alkenyl-, -C2- _C6 alkynyl-, -C1- _C2 alkyl-OC1 _{- C2} alkyl- or -C1 _{- C2} alkyl-NRa ^- _C1 - _C2 alkyl- group, the substituted _-C2 - _C6 alkyl- group being substituted by 1-2 substituents each independently selected _from the group consisting of halogen, hydroxy _, -OP(O)(OH) ₂ , -OP ₍ O)(R ^I R ^II ) ₂ , amino, (C1- _C4 alkyl)amino-, ( _C1 - _C4 alkyl)( _{C1 -C4 alkyl} )amino- _{, halo(C1 -} C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, q is 1, ^RA1 and ^RA2 are each independently _-CH2- or -O-, and A is a _-C2 - _C4 alkyl-, _-C2 - _C4 alkenyl- or _-C2 - _C4 alkynyl- group.

In selected _embodiments , q is 1, ^RA1 and ^RA2 are each -O-, _{and A is -CH2CH2CH2- ,} wherein A together with ^RA1 and ^RA2 form _{a -OCH2CH2CH2O-} group.

In another embodiment, q is 1, ^RA1 and ^RA2 are each -O-, and A is _-CH2 -phenyl- _CH2- , wherein A together with ^RA1 and ^RA2 form a _-OCH2 -phenyl- _CH2O- group. In a specific embodiment, q is 1, A together with ^RA1 and ^RA2 form a _-OCH2 -phenyl- _CH2O- group, wherein the _-OCH2- group is located at the 1,4 position on the phenyl ring portion.

The length of the linking group defined herein represents the lowest number of atoms in the direct chain consisting of ^-RA1 - ^ARA2- and/or ^-RB1 - ^BRB2- and/or ^-RC1 - ^CRC2- . For example, when B is an optionally substituted phenyl group, the linking group ^-RB1 - ^BRB2- can be represented as -( _CH2 )-phenyl-( _CH2 )-. This linking group is characterized as a 4-membered linking group when the two -( _CH2 )- moieties are located on adjacent carbon atoms of the benzene ring (1,2 substituted phenyl). In another embodiment, this linking group is characterized as a 6-membered linking group when the two -( _CH2 )- moieties are substituted at opposite positions on the benzene ring (1,4 substituted phenyl). It should be understood that any alkyl, alkenyl or alkynyl group or moiety of A, B or C is a straight or branched -alkyl, alkenyl or alkynyl group or moiety. For example, the ^-RB1 - ^BRB2- linking group, wherein B is _-C1 - _C10 alkyl-, may contain an 8-membered linking group having one ( _C1 - _C4 alkyl) branching group or 2-4 ( _C1 - _C3 alkyl) branching groups, such as 4 branched methyl groups (2 gem-dimethyl groups) or 2 branched methyl groups.

In one embodiment of the compounds of the present invention, r is 1, and ^RB1 and ^RB2 are each independently _-CH2- , and B together with ^RB1 and ^RB2 form a linking group, wherein B is a bond or B is -halo( _C1 - _C10 alkyl)-, optionally substituted- _C1 - _C10 alkyl-, optionally substituted- _C2 - _C10 alkenyl-, optionally substituted- _C2 - _C10 alkynyl-, optionally substituted- _C1 - _C6 alkyl- _OC1 - _C6 alkyl-, optionally substituted- _C1 - _C6 alkyl- ^NRa - _C1 - _C6 alkyl-, optionally substituted _C3 - _C6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl, optionally substituted- _C1 - _C4 alkyl-( _C3 - _C6 cycloalkyl)-C1- _C4 alkyl-, optionally substituted- _{C1 -} C6 -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl-, or optionally substituted -C ₁ -C ₄ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₄ alkyl-,

wherein the alkyl portion of the optionally substituted -C ₁ -C ₁₀ alkyl-, optionally substituted -C ₂ -C ₁₀ alkenyl-, optionally substituted -C ₂ -C ₁₀ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₄ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-phenyl-C ₁ -C ₄ alkyl-, optionally substituted -C ₁ -C ₄ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₄ alkyl- or optionally substituted -C ₁ -C ₄ alkyl-(5-6 membered heteroaryl-C ₁ -C ₄ alkyl)- is optionally substituted with 1-C 4 alkyl- wherein the at least one aryl group is substituted with 4 substituents each independently selected from the group consisting of -C ₁ -C ₄ alkyl, halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C ₆ cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl moiety of the optionally substituted C 3 -C 6 cycloalkyl, optionally substituted phenyl, optionally substituted 4-6-membered heterocycloalkyl, optionally substituted 5-6-membered heteroaryl, optionally substituted _{-C 1} -C ₄ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C _{4 alkyl-, optionally substituted -C 1 -C 4 alkyl - phenyl} -C ₁ -C ₄ alkyl-, optionally substituted -C 1 -C ₄ alkyl-(4-6-membered heterocycloalkyl)-C _{1 -C 4} alkyl- or optionally substituted -C ₁ -C ₄ alkyl-(5-6-membered heteroaryl)-C ₁ -C ₄ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the following: halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-.

In one embodiment of the compounds of the present invention, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B, ^RB1 and ^RB2, together form a 2-6 membered linking group. In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B, ^RB1 and ^RB2, together form a 3-6 membered linking group. In yet another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B, ^RB1 and ^RB2, together form a 4-5 membered linking group.

In one embodiment of the compounds of the present invention, ^r is 1, ^RB1 and ^RB2 are each independently ^-CRdRf- , and B, ^RB1 and ^RB2, together form a 2-6 membered linking group. In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B, ^RB1 and ^RB2, together form a 3-6 membered linking group. In yet another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B, ^RB1 and ^RB2, together form a 4-5 membered linking group.

In one embodiment, B is a bond.

In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B is a substituted _-C1 - _C10 alkyl-group or an unsubstituted _-C1 - _C10 alkyl-, -C2- _C10 alkenyl-, -C2- _C10 alkynyl-, _-C1 - _C6 alkyl- _OC1 - _C6 alkyl- or -C1 _{- C6} alkyl- ^NRa - _{C1 - C6} alkyl-group, the substituted _-C1 - _C10 alkyl-group being substituted by 1-4 substituents each independently selected from the group consisting of _-C1 - _C4 alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, ( _C1 - _C6 alkyl)amino-, (C1- _C6 alkyl)( _C1 - _C6 alkyl)amino-, halo( _{C1 - C alkyl} )-, -C ₁ -C ₆ alkyl), halo(C ₁ -C ₆ alkyl), halo(C ₁ -C 6 alkyl)-, C ₁ -C ₄ alkoxy-, hydroxy-(C 2 -C ₄ alkoxy)-, C ₁ -C 4 alkoxy-(C 1 -C ₄ alkoxy)-, -NHCO(C ₁ -C ₄ alkyl), optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl or 5-6 membered heteroaryl is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of -C ₁ -C ₄ alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl)amino-, halo(C ₁ -C 6 alkyl), halo(C ₁ -C ₆ alkyl), In _{some embodiments} , the _{present invention} comprises hydroxy-(C ₂ -C ₄ alkoxy)-, hydroxy-(C ₂ -C ₄ alkoxy)-, and hydroxy-(C ₁ -C ₄ alkoxy)-.

In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B is a substituted _-C1 - _C10 alkyl-group or an unsubstituted _-C1 - _C10 alkyl-, -C2- _C10 alkenyl-, -C2- _C10 alkynyl-, _-C1 - _C6 alkyl- _OC1 - _C6 alkyl- or _-C1 - _C6 alkyl- ^NRa - _{C1 - C6} alkyl-group, the substituted _-C1 - _C10 alkyl-group being substituted by 1-4 substituents each independently selected from the group consisting of _-C1 - _C4 alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, ( _C1 - _C4 alkyl)amino-, (C1- _C4 alkyl)( _C1 - _C4 alkyl)amino-, halo( _{C1 - C 4} alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B is a substituted _-C1 - _C8 alkyl-group or an unsubstituted _-C1 - _C8 alkyl-, _{-C2- C8} alkenyl-, -C2- _C8 alkynyl-, _-C1 - _C4 alkyl- _OC1 - _C4 alkyl- or -C1- _C4 alkyl- ^NRa - _{C1 - C4} alkyl-group, the substituted _-C1 - _C8 alkyl-group being substituted by 1-4 substituents each independently selected from the group consisting of _-C1 - _C4 alkyl, halogen, hydroxy, _-OP (O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, ( _C1 - _C4 alkyl)amino-, (C1- _C4 alkyl)( _C1 -C4 alkyl)amino-, halo( _C1 - _{C4 alkyl} ) ₄ alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B is a substituted _-C1 - _C6 alkyl-group or an unsubstituted _-C1 - _C6 alkyl-, -C2 _-C6 alkenyl-, -C2- _C6 alkynyl-, -C1- _C2 alkyl- _OC1 - _C2 alkyl- or -C1 _{- C2} alkyl- ^NRa - _{C1 - C2} alkyl-group, the substituted _-C1 - _C6 alkyl-group being substituted by 1-4 substituents each independently selected from the group consisting of _-C1 - _C4 alkyl, halogen _{, hydroxy, -OP(O)(OH)2 ,} -OP(O)(R ^I R ^II ) ₂ , amino, ( _C1 - _C4 alkyl)amino-, (C1- _C4 alkyl)( _C1 - _C4 alkyl)amino-, halo( _{C1 - C 4} alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B is a substituted _-C2 - _{C4 alkyl-group or an unsubstituted -C2-C4 alkyl-, -C2-C4 alkenyl-, -C2-C4 alkynyl-, -C1 alkyl-OC1 alkyl- or -C1 alkyl-NRa-C1 alkyl-group, wherein the substituted -C2-C4 alkyl - group is substituted with 1-4 substituents each independently selected} from the group consisting of _-C1 - _C4 alkyl, halogen, hydroxy, -OP(O)( _OH ) ₂ , -OP(O) ⁽ R ^I R ^II ) ₂ , amino, ( _C1 - _C4 alkyl)amino-, ( _C1 - _C4 alkyl)( _C1 - _C4 alkyl)amino-, halo( _C1 - _C4 alkyl), halo( _C1 - _C4 alkoxy)- and C1-C4 alkyl-. ₁ -C ₄ alkoxy-.

In one embodiment, r is 1, ^RB1 and ^RB2 are each independently ^{-CRdRf , Rd} and ^Rf are H or methyl, and B is -CH=CH-, -CH( _CH3 )=CH( _CH3 )-, _-CH2CH2- , _{-CH(OH)CH(OH)-, -CH(CH3)CH(CH3 ) -} , _-CF2 - _CF2- , or _-CH2CH2CH2- . In ^{one embodiment, r is 1, RB1 and RB2 are each independently -CRdRf, Rd and Rf are} _H ^{or methyl ,} and ^B is -CH= _CH- , -CH2CH2-, -CH( _OH )CH(OH ₎ -. In these embodiments, _r is 1, and B, together with ^RB1 and ^RB2, forms a _-CH2CH = _CHCH2- , _{-CH2CH2CH2CH2- , -CH2CH} ( _OH )CH(OH) _CH2- , or _-CH2CH2N ( _CH3 ) _CH2CH2- group. In these embodiments, r is 1, and B _, together with ^RB1 and ^RB2, _{forms -CH2CH = CHCH2-} . In one embodiment, r is 1, and B, together with ^RB1 and ^RB2, _{forms -CH2CH2CH2CH2- .}

In one embodiment of the compounds of the present invention, s is 1, and R ^C1 and R ^C2 are each independently -CH ₂ -, and C together with R ^C1 and R ^C2 form a linking group, wherein C is -halo(C ₁ -C ₁₂ alkyl)-, optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C 6 ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C ₆ alkyl-,

wherein the optionally substituted -C ₁ -C ₁₂ alkyl-, optionally substituted -C ₂ -C ₁₂ alkenyl-, optionally substituted -C ₂ -C ₁₂ alkynyl-, optionally substituted -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-NR ^a -C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(C ₃ -C ₆ cycloalkyl)-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C ₁ -C ₆ alkyl-(4-6 membered heterocycloalkyl)-C ₁ -C ₆ alkyl- or optionally substituted -C ₁ -C ₆ alkyl-(5-6 membered heteroaryl)-C ₁ -C The alkyl portion of the ₆ -alkyl- is optionally substituted with 1 or 2 substituents each independently selected from the group consisting of halogen, halo(C ₁ -C ₄ alkyl), -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^d , -OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c and -NR ^d SO ₂ R ^c ,

and

The C ₃ -C _{6 cycloalkyl, phenyl, 4-6-membered heterocycloalkyl or 5-6-membered heteroaryl portion of the optionally substituted -C 1 -C 6 alkyl-(C 3 -C 6 cycloalkyl)-C 1 -C 6 alkyl- , optionally} substituted -C ₁ -C 6 alkyl-phenyl-C ₁ -C ₆ alkyl-, optionally substituted -C _{1 -C 6 alkyl-(4-6-membered heterocycloalkyl)-C 1 -C 6 alkyl- or optionally substituted -C 1 -C 6 alkyl- ( 5-6 -} membered heteroaryl)-C ₁ -C ₆ alkyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C 6 _{R I R II ) 2 and C 1 -C 4 alkoxy- ( C 1 -C 4} ^{alkoxy )} _{- .}

In one embodiment of the compounds of the present invention, s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C together with R ^C1 and R ^C2 forms a 4-8 membered linking group. In another embodiment, s is 1, and C together with R ^C1 and R ^C2 forms a 4-6 membered linking group. In yet another embodiment, s is 1, and C together with R ^C1 and R ^C2 forms a 5-membered linking group.

In another embodiment, s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C is a substituted -C ₂ -C ₁₀ alkyl- group or an unsubstituted -C ₂ -C ₁₀ alkyl-, -C 2 -C ₁₀ alkenyl-, -C ₂ -C ₁₀ alkynyl-, -C ₁ -C ₄ alkyl-OC ₁ -C ₄ alkyl- or -C ₁ -C ₄ alkyl-NR ^a -C ₁ -C ₄ alkyl- group, the substituted -C ₂ -C ₁₀ alkyl- group being substituted by _1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, halo(C ₁ -C ₄ alkyl), halo(C ₁ -C ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C is a substituted -C ₂ -C ₈ alkyl- group or an unsubstituted -C ₂ -C ₈ alkyl-, -C 2 -C ₈ alkenyl-, -C ₂ -C ₈ alkynyl-, -C ₁ -C ₂ alkyl-OC ₁ -C ₂ alkyl- or -C ₁ -C 2 alkyl-NR ^a -C ₁ -C ₂ alkyl- group, the substituted -C ₂ -C ₈ alkyl- group being substituted by _1-2 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C _{1 -C 4 alkyl)amino-, (C 1 -C 4 alkyl} )(C ₁ -C ₄ alkyl)amino-, halo(C 1 -C ₄ alkyl), halo(C _{1 -C 4} alkyl), ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C is a substituted -C ₂ -C ₆ alkyl- group or an unsubstituted -C ₂ -C ₆ alkyl-, -C 2 -C ₆ alkenyl-, -C ₂ -C ₆ alkynyl-, -C ₁ -C ₂ alkyl-OC ₁ -C ₂ alkyl- or -C ₁ -C 2 alkyl-NR ^a -C ₁ -C ₂ alkyl- group, the substituted -C ₂ -C ₆ alkyl- group being substituted by _1-2 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C _{1 -C 4 alkyl)amino-, (C 1 -C 4 alkyl} )(C ₁ -C ₄ alkyl)amino-, halo(C 1 -C ₄ alkyl), halo(C _{1 -C 4} alkyl), ₄ alkoxy)- and C ₁ -C ₄ alkoxy-.

In another embodiment, s is 1, R ^C1 and R ^C2 are each independently -CH ₂ -, and C is a -C ₂ -C ₄ alkyl-, -C ₂ -C ₄ alkenyl- or -C ₂ -C ₄ alkynyl- group.

In selected embodiments, s is 1, ^RC1 and ^RC2 are each _{independently -CH2- , and C is -CH2CH2CH2- , wherein} C together with ^RC1 _and ^RC2 form _{a -CH2CH2CH2CH2CH2- group} .

In one embodiment of the compounds of the present invention, ^R4 and ^R6 are each independently selected from H, halogen, halo( _C1 - _C6 alkyl), halo( _C1 - _C6 alkoxy)-, hydroxy, -OP(O)(OH) ₂ , -OP(O)( ^{R I} R ^II ) ₂ , _-NH2 , -NRcRc ^{, -NRcRd} , ^-CORc , _-CO2Rc , -N( ^Rd ) ^CORc , ^-N ( ^{Rd )} _SO2Rc , -N( ^Rg ) _SO2 (C1-C2 alkyl)-N( ^Rh )( ^Rf ), -N( ^{Rg )} CO( _C1 - _C2 alkyl)-N( ^Rh )( ^Rf ), optionally substituted ( _C1 - _C6 alkyl), optionally substituted ( _C1 - _C6 alkyl)oxy-, optionally substituted ( _{C1 - C6} alkyl)oxy-, ₆ alkyl)amino- and optionally substituted (C ₁ -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-,

wherein the optionally substituted (C ₁ -C ₆ alkyl), optionally substituted (C ₁ -C ₆ alkyl)oxy-, optionally substituted (C ₁ -C ₆ alkyl)amino- and optionally substituted (C ₁ -C ₆ alkyl)(C ₁ -C ₄ alkyl)amino-(C ₁ -C ₆ alkyl) are optionally substituted by 1 to 4 substituents each independently selected from the group consisting of -OH, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -OR ^c , -NH ₂ , -NR ^c R ^c , -NR ^c R ^d , -CO ₂ H , -CO ₂ R ^c , OCOR ^c , -CO ₂ H , -CO ₂ R ^c , -SOR ^c , -SO ₂ R ^c , -CONH ₂ , -CONR ^c R ^d , -SO ₂ NH ₂ , -SO ₂ NR ^c R ^d , -OCONH ₂ , -OCONR ^c R ^d , -NR ^d COR ^c , -NR ^d SOR ^c , -NR ^d CO ₂ R ^c , -NR ^d SO ₂ R ^c , optionally substituted phenyl, optionally substituted 5-6 membered heterocycloalkyl, and optionally substituted 5-6 membered heteroaryl, wherein the optionally substituted phenyl, 5-6 membered heterocycloalkyl, or 5-6 membered heteroaryl is optionally substituted with 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, amino, (C ₁ -C ₄ alkyl)amino-, (C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl)amino-, C ₁ -C ₄ alkyl, halo(C ₁ -C ₄ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-, -COR ^d , -CON(R ^d )(R ^f ) and -CO ₂ R ^d .

In one embodiment, ^R4 and ^R6 are each H.

In one embodiment, R ³ and R ⁵ are independently selected from -CO-N(R ^d )(R ^f ), and each R ^d and R ^f is independently H or C ₁ -C ₃ alkyl.

In one embodiment, R ³ and R ⁵ are CONH ₂ .

In one embodiment of the compounds of the present invention, ^R14 is optionally substituted _C1 - _C4 ^alkyl , wherein said optionally substituted _C1 - _C4 alkyl is optionally substituted with a substituent selected from the group consisting of ^{-ORc , -NRcRd} , _-CO2Rc ^{, -CONRcRd ,} _-SO2NRcRd ^{and -OCONRcRd .}

In one embodiment of the compounds of the present invention, R ¹⁶ is H, halogen or C ₁ -C ₄ alkyl.

In one embodiment of the compounds of the present invention, R ¹⁵ and R ¹⁷ are each independently H, cyclopropyl or C ₁ -C ₄ alkyl.

In one embodiment of the compounds of the present invention, R ¹⁴ , R ¹⁵ , R ¹⁶ and R ¹⁷ are each independently H or C ₁ -C ₄ alkyl.

In one embodiment of the present invention, R ¹⁶ is H.

In another embodiment, R ¹⁴ , R ¹⁵ and R ¹⁷ are each independently C ₁ -C ₄ alkyl.

In another embodiment, R ¹⁴ , R ¹⁵ and R ¹⁷ are each independently C ₁ -C ₃ alkyl, specifically methyl or ethyl. In a selected embodiment, R ¹⁴ is ethyl.

In another embodiment, R ¹⁵ and R ¹⁷ are each methyl.

In one embodiment of the compounds of the present invention, ^Ra is H, ^-Rc , _-CORc , ^-CO2H , _-CO2Rc ^{, -SORc , -SO2Rc} , _-CONH2 , _-CONRcRd ^{, -SO2NH2} _{, or} ^-SO2NRcRd _.

In another embodiment, ^Ra is H, _C1 - _C4 alkyl, -CO( _C1 - _C4 alkyl), -CO( _C1 - _C4

-CO(C ₁ -C ₄ alkyl)-OH, -CO(C ₁ -C ₄ alkyl)-O-(C ₁ -C ₄ alkyl), -CO(C ₁ -C ₄ alkyl)-NH ₂ , -CO(C ₁ -C _{4 alkyl)-NH(C 1 -C 4} alkyl), or -CO(C ₁ -C ₄ alkyl)-N(C ₁ -C ₄ alkyl)(C ₁ -C ₄ alkyl).

In one embodiment of the compounds of the present invention, ^Rx and ^Ry are each independently methyl or ethyl. In one embodiment of the compounds of the present invention, ^Rx and ^Ry are both methyl. In one embodiment of the compounds of the present invention, the compound of formula (I), wherein one of ^Rx and ^Ry is methyl and the other is H.

One embodiment of the present invention relates to compounds of formula (I), wherein:

q + r + s = 1 or 2;

q is 0, and ^RA1 and ^RA2 are independently selected _from H _{, -OCH2CH2CH2OH} and _-OCH3 ; or

q is 1, ^RA1 and ^RA2 are each -O-, and A is -CH ₂ CH ₂ CH ₂ -;

r is 0, and ^RB1 and ^RB2 are each H; or

r is 1, ^RB1 and ^RB2 are each independently _-CH2- , and B is -CH=CH- _{, -CH2CH2- ,} -CH(OH)CH(OH)-, or _-CH2N ( _CH3 ) _CH2- ;

s is 0, R ^C1 is methyl and R ^C2 is ethyl; or

s is 1, ^RC1 and ^RC2 are each independently -CH ₂ -, and C is -CH ₂ CH ₂ CH ₂ -;

R ³ and R ⁵ are each -CONH ₂ ;

^R4 and ^R6 are each H;

R ¹⁴ is ethyl;

R ¹⁵ is methyl;

^R16 is H;

R ¹⁷ is a methyl group,

or a salt thereof, particularly a pharmaceutically acceptable salt.

In one embodiment of the compounds of the present invention,

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl;

One of R ^A1 and R ^A2 is H and the other of R ^A1 and R ^A2 is optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted by 1 to 2 substituents each independently selected from the following: hydroxyl and optionally substituted phenyl,

wherein the optionally substituted phenyl is optionally substituted by 1-2 substituents each independently selected from (C ₁ -C ₄ alkyl)oxy-;

^R3 and ^R5 are both -CO- _NH2 ; and

B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: fluorine and C _1-2 alkyl; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

Or both ^RX and ^Ry are independently _C1 - _C4 alkyl.

In one embodiment of the present invention, the compound of the present invention is of formula (I-B')

in

R ³ and R ⁵ are each independently -CON(R ^d )(R ^f ), or one of R ³ and R ⁵ is -CON(R ^d )(R ^f ), and the other of R ³ and R ⁵ is H or -CO ₂ (R ^c );

R ^c is C ₁ -C ₄ alkyl;

^RB1 and ^RB2 are each independently ^{-CRdRf- ;}

B is -halo(C ₁ -C ₅ alkyl), unsubstituted -C ₁ -C ₅ alkyl or unsubstituted -C ₂ -C ₅ alkenyl-;

R ^A2 and R ^A1 are each independently H, halogen, hydroxy, optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the C ₁ -C ₆ alkyl group of the optionally substituted (C ₁ -C _{6 alkyl) or optionally substituted (C 1 -C 6 alkyl )} oxy- is optionally substituted by 1-4 substituents each independently selected from the following: hydroxyl, C ₁ -C ₄ alkoxy, -N( ^Re )( ^Rf ), _-CO2 ( ^Rf ), optionally substituted phenyl and optionally substituted 5-6 membered heterocycloalkyl; wherein the optionally substituted phenyl or 5-6 membered heterocycloalkyl is optionally substituted by 1-4 substituents each independently selected from the following:

C ₁ -C ₄ alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl)amino-, halo(C ₁ -C ₆ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

Each R ^d is independently H or C ₁ -C ₄ alkyl;

R ^e is selected from H, (C ₁ -C ₄ alkyl), -CO(C ₁ -C ₄ alkyl), -OCO(C ₁ -C ₄ alkyl) or -CO ₂ (C ₁ -C ₄ alkyl);

Each R ^f is H or (C ₁ -C ₄ alkyl);

^R4 and ^R6 are H;

R ¹⁴ is C ₁ -C ₄ alkyl;

R ^C1 is H or C ₁ -C ₄ alkyl;

R ^C2 is C ₁ -C ₄ alkyl;

R ¹⁵ is H or C _{1 -} C ₄ alkyl;

R ¹⁶ is H or C _{1 -} C ₄ alkyl;

R ¹⁷ is H or C _{1 -C 4} alkyl; and

R ^I and R ^II are independently (C ₁ -C ₆ alkyl)oxy- at each occurrence,

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof,

or a salt thereof.

In one embodiment of the present invention, the compound of the present invention is of formula (I-b'),

in

B is -halo(C ₁ -C ₅ alkyl), unsubstituted -C ₁ -C ₅ alkyl, or unsubstituted -C ₂ -C ₅ alkenyl-;

^RA2 and ^RA1 are each independently H, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the C ₁ -C ₆ alkyl group of the optionally substituted (C ₁ -C _{6 alkyl) or optionally substituted (C 1 -C 6 alkyl )} oxy- is optionally substituted by 1-4 substituents each independently selected from the group consisting of hydroxy, C ₁ -C ₄ alkoxy, -N( ^Re )( ^Rf ), _-CO2 ( ^Rf ), optionally substituted phenyl, and optionally substituted 5-6 membered heterocycloalkyl; and wherein the optionally substituted phenyl or 5-6 membered heterocycloalkyl group is optionally substituted by 1-4 substituents each independently selected from the group consisting of C ₁ -C ₄ alkyl, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C ₆ alkyl)amino-, halo(C ₁ -C ₆ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -(C 1 -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R _I R _II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , -(C ₁ -C ₆ alkyl)-NH ₂ , -C ₁ -C ₄ alkyl-(C ₁ -C ₄ alkoxy) and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

R ^e is selected from H, (C ₁ -C ₄ alkyl), -CO(C ₁ -C ₄ alkyl), -OCO(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-NH ₂ , -(C ₁ -C ₄ alkyl)C ₁ -C ₄ alkoxy or -CO ₂ (C ₁ -C ₄ alkyl),

Each R ^f is H or (C ₁ -C ₄ alkyl);

R ¹⁴ is C ₁ -C ₄ alkyl;

R ^C2 is C ₁ -C ₄ alkyl;

R ¹⁵ is C _{1 -C 4} alkyl; and

R ¹⁷ is C _{1 -} C ₄ alkyl;

R ^I and R ^II are independently (C ₁ -C ₆ alkyl)oxy- at each occurrence,

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof,

or a salt thereof.

In one embodiment, the compound of formula (I-B') or (I-b'), wherein ^RA2 and ^RA1 are each independently H, halogen, optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1- _C6 alkyl of the optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _{C1 - C6} alkyl)oxy- is optionally substituted with 1-4 substituents each independently selected from the group consisting of hydroxy, -OP(O)(OH) ₂ , -OP(O)( ^{RIRII )} ₂ , -N( ^Re )( ^Rf ), _C1 - _C4 alkoxy, phenyl, an optionally substituted 5-6 membered heterocycloalkyl containing at least one nitrogen or oxygen as a ring member, and each ^Re is independently selected from H, ( _C1 - _C4 alkyl), -( _C1 - _C4 alkyl) _-NH2 or -( _C1 - _C4 alkyl) _C1 -C ₄ alkoxy and each R ^f is independently H or (C ₁ -C ₄ alkyl).

In one embodiment, the compound of formula (I-B') or (I-b'), wherein ^RA2 and ^RA1 are each independently H, halogen, optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1- _C6 alkyl of the optionally substituted ( _C1 - _{C6 alkyl), optionally substituted (C1-C6 alkyl ) oxy-} is optionally substituted with 1-4 substituents each independently selected from the following substituents: hydroxy, -N( ^Re )( ^Rf ), _C1 - _C4 alkoxy, phenyl, optionally substituted 5-6 membered heterocycloalkyl containing at least one nitrogen or oxygen as a ring member, and ^Re and ^Rf are each independently H or ( _C1 - _C4 alkyl).

In one embodiment, a compound of formula (I-B') or (I-b'), wherein at least one of ^RA2 or ^RA1 is independently H, halogen, optionally substituted ( _C1 - _C6 alkyl), or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1- _{C6 alkyl of the optionally substituted (C1-C6 alkyl), optionally substituted (C1-C6 alkyl ) oxy- is optionally} substituted with 1-4 substituents each independently selected from the group consisting of -N( ^Re )( ^Rf ), tetrahydropyran, pyrrolidinyl, piperazinyl, piperidinyl, and morpholinyl, each ^Re is independently selected from H, ( _C1 - _C4 alkyl), -( _C1 - _C4 alkyl)-NH2 _, or -( _C1 - _C4 alkyl) _C1 - _C4 alkoxy, and each ^Rf is independently H or ( _C1 - _C4 alkyl).

In one embodiment, a compound of formula (I-B') or (I-b'), wherein at least one of ^RA2 or ^RA1 is each independently H, halogen, optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1- _C6 alkyl of the optionally substituted ( _C1 - _C6 alkyl) or optionally substituted ( _C1 - _{C6 alkyl} )oxy- is optionally substituted with 1-4 substituents each independently selected from the following substituents: -N( ^Re )( ^Rf ), tetrahydropyran, pyrrolidinyl, piperazinyl, piperidinyl or morpholinyl, and ^Re and ^Rf are each independently H or ( _C1 - _C4 alkyl).

In one embodiment, the compound of formula (I-B') or (I-b'), wherein ^Rx and ^Ry are each independently methyl or ethyl. In another embodiment, the compound of formula (I-B') or (I-b'), wherein both ^Rx and ^Ry are methyl. In another embodiment, the compound of formula (I-B') or (I-b'), wherein one of ^Rx and ^Ry is methyl and the other is H.

In one embodiment, the compound of formula (I-B') or (I-b'), wherein

B is unsubstituted -C ₁ -C ₅ alkyl or unsubstituted -C ₂ -C ₅ alkenyl-;

R ^A2 and R ^A1 are each independently H, halogen, optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the C ₁ -C ₆ alkyl group of the optionally substituted (C ₁ -C _{6 alkyl) or optionally substituted (C 1 -C 6 alkyl )} oxy- is optionally substituted by 1-2 substituents each independently selected from the following: hydroxyl, C ₁ -C ₄ alkoxy, -N( ^Re )( ^Rf ), _-CO2 ( ^Rf ), unsubstituted phenyl and unsubstituted 5-6 membered heterocycloalkyl,

R ^e is H, (C ₁ -C ₄ alkyl), -CO(C ₁ -C ₄ alkyl), -OCO(C ₁ -C ₄ alkyl) or -CO ₂ (C ₁ -C ₄ alkyl),

R ^f at each occurrence is H or (C ₁ -C ₄ alkyl);

R ¹⁴ is C ₁ -C ₄ alkyl;

R ^C2 is C ₁ -C ₄ alkyl;

R ¹⁵ is C _{1 -C 4} alkyl; and

R ¹⁷ is C _{1 -} C ₄ alkyl;

R ^I and R ^II are independently (C ₁ -C ₆ alkyl)oxy- at each occurrence,

At least one of ^RX or ^Ry is independently C ₁ -C ₂ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₂ alkyl;

or a tautomer thereof,

or a salt thereof.

In one embodiment, the compound of formula (I-b') wherein

B is unsubstituted -C ₂ -C ₅ alkenyl-;

R ^A2 and R ^A1 are each independently H, optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-C ₁ -C ₆ alkyl is optionally substituted by 1 substituent independently selected from the following: hydroxyl, C ₁ -C ₄ alkoxy, unsubstituted 5-6 membered heterocycloalkyl,

R ¹⁴ is C ₁ -C ₄ alkyl;

R ^C2 is C ₁ -C ₄ alkyl;

R ¹⁵ is C _{1 -C 4} alkyl; and

R ¹⁷ is C _{1 -} C ₄ alkyl;

At least one of ^RX or ^Ry is independently C ₁ -C ₂ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₂ alkyl;

or a tautomer thereof,

or a salt thereof.

In one embodiment, the compound of formula (I-b') wherein

B is an unsubstituted vinyl group;

R ^A2 and R ^A1 are each independently H or an optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the C ₁ -C ₆ alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted with a substituent selected from hydroxyl or unsubstituted morpholinyl;

R ¹⁴ is methyl or ethyl;

R ^C2 is methyl or ethyl;

^R15 is methyl or ethyl; and

R ¹⁷ is methyl or ethyl;

At least one of ^RX or ^Ry is independently C ₁ -C ₂ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₂ alkyl;

or a tautomer thereof,

or a salt thereof.

In one embodiment, the compound of formula (I-1),

in

P is an integer from 1 to 6;

^RA and ^RB are independently H or _C1 - _C4 alkyl;

or N, ^RA and ^RB form an optionally substituted 5- or 6-membered heterocyclic ring,

wherein the heterocyclic ring is selected from morpholinyl, piperidinyl, piperazinyl and pyrrolidinyl, and

The heterocycle is optionally substituted with one or two substituents independently selected from the group consisting of hydroxy and C ₁ -C ₃ alkyl optionally substituted with one or two substituents which are hydroxy or C ₁ -C ₃ alkoxy;

At least one of ^RX or ^Ry is independently _C1 - _C2 alkyl and the other is H;

or R ^x and R ^y are both independently C ₁ -C ₂ alkyl;

or a tautomer thereof,

or a salt thereof.

In one embodiment, the compound of formula (I-1), wherein ^Rx and ^Ry are each independently methyl or ethyl. In another embodiment, the compound of formula (I-1), wherein both ^Rx and ^Ry are methyl. In another embodiment, the compound of formula (I-1), wherein one of ^Rx and ^Ry is methyl and the other is H.

In one embodiment, the compound of the invention has the formula (I-bc)

in

R ^C1 and R ^C2 are each independently -CH ₂ -,

C is -halo(C ₁ -C ₅ alkyl), unsubstituted -C ₁ -C ₅ alkyl or unsubstituted -C ₂ -C ₅ alkenyl-;

^RB1 and ^RB2 are each independently ^{-CRdRf- ;}

B is -halo(C ₁ -C ₅ alkyl), unsubstituted -C ₁ -C ₅ alkyl or unsubstituted -C ₂ -C ₅ alkenyl-;

^RA2 and ^RA1 are each independently H, halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the C ₁ -C ₆ alkyl group of the optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy- is optionally substituted by 1-4 substituents each independently selected from the group consisting of hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , C ₁ -C ₄ alkoxy, -N( ^Re )(R ^f ), -CO ₂ (R ^f ), optionally substituted phenyl, and optionally substituted 5-6 membered heterocycloalkyl; wherein the optionally substituted phenyl or 5-6 membered heterocycloalkyl is optionally substituted by 1-4 substituents each independently selected from the group consisting of halogen, hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , amino, (C ₁ -C ₆ alkyl)amino-, (C ₁ -C ₆ alkyl)(C ₁ -C 6 alkyl)amino-, halo(C ₁ -C ₄ alkoxy) ₆ alkyl), hydroxy-(C ₁ -C ₄ alkyl)-, -(C ₁ -C ₄ alkyl)-OP(O)(OH) ₂ , -(C ₁ -C ₄ alkyl)-OP(O)(R ^I R ^II ) ₂ , halo(C ₁ -C ₄ alkoxy)-, C ₁ -C ₄ alkoxy-, hydroxy-(C ₂ -C ₄ alkoxy)-, -(C ₂ -C ₄ alkoxy)-OP(O)(OH) ₂ , -(C ₂ -C ₄ alkoxy)-OP(O)(R ^I R ^II ) ₂ , -(C ₁ -C ₆ alkyl)-NH ₂ and C ₁ -C ₄ alkoxy-(C ₁ -C ₄ alkoxy)-;

Each R ^d is independently H or C ₁ -C ₄ alkyl;

R ^e is selected from H, (C ₁ -C ₄ alkyl), -CO(C ₁ -C ₄ alkyl), -OCO(C ₁ -C ₄ alkyl), -(C ₁ -C ₄ alkyl)-NH ₂ , -(C ₁ -C ₄ alkyl)C ₁ -C ₄ alkoxy or -CO ₂ (C ₁ -C ₄ alkyl),

Each R ^f is H or (C ₁ -C ₄ alkyl);

^R6 is H;

R ¹⁴ is C ₁ -C ₄ alkyl;

R ¹⁵ is C _{1 -} C ₄ alkyl;

R ¹⁶ is C _{1 -} C ₄ alkyl;

R ¹⁷ is C _{1 -C 4} alkyl; and

R ^I and R ^II are independently (C ₁ -C ₆ alkyl)oxy- at each occurrence,

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof;

or a salt thereof, in particular a pharmaceutically acceptable salt thereof.

In one embodiment, a compound of formula (I-bc) wherein ^R and ^R are each independently H, halogen, optionally substituted (C ₁ -C ₆ alkyl) or optionally substituted (C ₁ -C ₆ alkyl)oxy-, and the C ₁ -C ₆ alkyl of the optionally substituted (C ₁ -C ₆ alkyl)oxy- is optionally substituted with 1-4 substituents each independently selected from the group consisting of hydroxy, -OP(O)(OH) ₂ , -OP(O)(R ^I R ^II ) ₂ , -N( ^Re )(R ^f ), C _{1 -C 4} alkoxy, phenyl, an optionally substituted 5-6 membered heterocycloalkyl containing at least one nitrogen or oxygen as a ring member, each ^Re is independently selected from _H , C ₁ -C ₄ alkyl, -(C ₁ -C ₄ alkyl)-NH ₂ or -(C ₁ -C ₄ alkyl)C ₁ -C ₄ alkoxy, and each R ^f is independently H or (C ₁ -C ₄ alkyl).

In one embodiment, a compound of formula (I-bc) wherein at least one of ^RA2 or ^RA1 is independently H, halogen, optionally substituted ( _C1 - _C6 alkyl), or optionally substituted ( _C1 - _C6 alkyl)oxy-, and the C1- _C6 alkyl of the optionally substituted ( _C1 - _C6 alkyl), optionally substituted ( _{C1 - C6} alkyl)oxy- is optionally substituted with 1-4 substituents each independently selected from the group consisting of -N( ^Re )( ^Rf ), tetrahydropyran, pyrrolidinyl, piperazinyl, piperidinyl, and morpholinyl, each ^Re is independently selected from H, _C1 - _C4 alkyl, -( _C1 - _C4 alkyl) _-NH2 , or -( _C1 - _C4 alkyl) _C1 - _C4 alkoxy, and each ^Rf is independently H or _C1 - _C4 alkyl.

In one embodiment, the compound of formula I-2,

in

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently C ₁ -C ₃ alkyl;

R ^A1 and R ^A2 are independently H, hydroxy, COOH or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted by 1 to 4 substituents each independently selected from the group consisting of hydroxy, -CO ₂ (R ^f ), -N(R ^e )(R ^f ), optionally substituted phenyl and optionally substituted 5-6 membered heterocycloalkyl,

wherein the optionally substituted phenyl or 5-6 membered heterocycloalkyl is optionally substituted by 1-4 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy- and C ₁ -C ₄ alkyl;

^R3 and ^R5 are each independently -CO-N( ^Rd )( ^Rf ),

Each of R ^d , ^Re and R ^f is independently H or C ₁ -C ₃ alkyl;

B is a substituted -C ₁ -C ₄ alkyl- or a substituted -C ₂ -C ₄ alkenyl-,

wherein the alkyl portion of the substituted -C ₁ -C ₄ alkyl- or substituted -C ₂ -C ₄ alkenyl- is substituted by 1 to 4 substituents independently selected from the group consisting of halogen, hydroxy, (C ₁ -C ₄ alkyl)oxy- and C _1-4 alkyl,

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof,

or a salt thereof (especially a pharmaceutically acceptable salt thereof).

In one embodiment, in the compound of formula (I-2), R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl.

In one embodiment, in the compound of formula (I-2), R ¹⁴ and R ^c2 are ethyl and R ¹⁵ and R ¹⁷ are methyl.

In one embodiment, in the compound of formula (I-2), R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are methyl.

In one embodiment, in the compound of formula (I-2), R ^A1 and R ^A2 are independently H, hydroxy or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted by 1 to 4 substituents each independently selected from the group consisting of hydroxyl, COOH and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1 to 4 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, in the compound of formula (I-2), one of ^RA1 and ^RA2 is H and the other of ^RA1 and ^RA2 is hydroxy or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted by 1 to 4 substituents each independently selected from the group consisting of hydroxyl, COOH and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1 to 4 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, in the compound of formula (I-2), one of ^RA1 and ^RA2 is H and the other of ^RA1 and ^RA2 is hydroxy or optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted by 1 to 2 substituents each independently selected from the following: hydroxyl and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1-2 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, in the compound of formula (I-2), both ^RA1 and ^RA2 are H.

In one embodiment, the compound of formula (I-2) wherein at least one of ^RA1 and ^RA2 is not H.

In one embodiment, in the compound of formula (I-2), ^RA1 and ^RA2 are each independently optionally substituted ( _C1 - _C4 alkyl)oxy-, wherein the alkyl group of the optionally substituted ( _C1 - _C4 alkyl)oxy- is optionally substituted with 1-2 hydroxy substituents.

In one embodiment, in the compound of formula (I-2), R ³ and R ⁵ are independently selected from -CO-N(R ^d )(R ^f ), and R ^d and R ^f are each independently H or C ₁ -C ₃ alkyl.

In one embodiment, in the compound of formula (I-2), R ³ and R ⁵ are -CO-NH ₂ .

In one embodiment, in the compound of formula (I-2), B is substituted -C ₁ -C ₄ alkyl- or substituted -C ₂ -C ₄ alkenyl-,

The alkyl portion of the substituted -C ₁ -C ₄ alkyl- or substituted -C ₂ -C ₄ alkenyl- is substituted with 1 to 4 substituents independently selected from the following: halogen and C _1-4 alkyl.

In one embodiment, in the compound of formula (I-2), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

The substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: halogen and C _1-4 alkyl.

In one embodiment of the compound of formula (I-2), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

The substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: fluorine and C _1-2 alkyl.

In one embodiment of the compound of formula (I-2), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 fluorine substituents.

In one embodiment of the compound of formula (I-2), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

The substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents each independently being a C _1-2 alkyl group.

In one embodiment of the compound of formula (I-2), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 hydroxy substituents.

In one embodiment of the compounds of formula (I-2), B is -CH ₂ -CH ₂ - substituted with 1-2 hydroxy substituents.

In one embodiment, the compound of formula (I-2), wherein ^Rx and ^Ry are each independently methyl or ethyl. In another embodiment, the compound of formula (I-2), wherein both ^Rx and ^Ry are methyl. In another embodiment, the compound of formula (I-2), wherein one of ^Rx and ^Ry is methyl and the other is H.

In one embodiment, the compound of formula (I-2), wherein

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl;

One of R ^A1 and R ^A2 is H and the other of R ^A1 and R ^A2 is optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted by 1 to 2 substituents independently selected from the following: hydroxyl and optionally substituted phenyl,

wherein the optionally substituted phenyl is optionally substituted by 1-2 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-;

^R3 and ^R5 are both -CO- _NH2 ; and

B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: fluorine and C _1-2 alkyl; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

Or both ^RX and ^Ry are independently _C1 - _C4 alkyl.

In one embodiment, the compound of formula (I-2), wherein

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl;

One of R ^A1 and R ^A2 is H and the other of R ^A1 and R ^A2 is optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted with 1-2 hydroxy substituents;

^R3 and ^R5 are both -CO- _NH2 ; and

B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: hydroxyl and C _1-2 alkyl; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

Or both ^RX and ^Ry are independently _C1 - _C4 alkyl.

In one embodiment, the compound of formula I-3,

in

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently C ₁ -C ₃ alkyl;

R ^A1 and R ^A2 are independently H, hydroxy, halogen, COOH or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted by 1 to 4 substituents each independently selected from the group consisting of hydroxy, -CO ₂ (R ^f ), -N( ^Re )(R ^f ), (C ₁ -C ₄ alkyl)oxy-, optionally substituted phenyl and optionally substituted 5-6 membered heterocycloalkyl,

wherein the optionally substituted phenyl or 5-6 membered heterocycloalkyl is optionally substituted by 1-4 substituents each independently selected from the group consisting of halogen, (C ₁ -C ₄ alkyl)oxy- and C ₁ -C ₄ alkyl;

R ³ and R ⁵ are each independently -CO-N(R ^d )(R ^f ), or one of R ³ and R ⁵ is -CO-N(R ^d )(R ^f ), and the other of R ³ and R ⁵ is H or CO ₂ (R ^C );

^R4 and ^R6 are each independently H or halogen,

Each of R ^c , R ^d , ^Re and R ^f is independently H or C ₁ -C ₄ alkyl;

B is an optionally substituted -C ₁ -C ₄ alkyl- or an optionally substituted -C ₂ -C ₄ alkenyl-,

wherein the alkyl portion of the optionally substituted -C ₁ -C ₄ alkyl- or optionally substituted -C ₂ -C ₄ alkenyl- is optionally substituted with 1 to 4 substituents each independently selected from the group consisting of halogen, hydroxy, (C ₁ -C ₄ alkyl)oxy- and C _1-4 alkyl; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

or R ^x and R ^y are both independently C ₁ -C ₄ alkyl;

or a tautomer thereof,

or a salt thereof (especially a pharmaceutically acceptable salt thereof).

In one embodiment, in the compound of formula (I-3), R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl.

In one embodiment, in the compound of formula (I-3), R ¹⁴ and R ^c2 are ethyl, and R ¹⁵ and R ¹⁷ are methyl.

In one embodiment, in the compound of formula (I-3), R ^A1 and R ^A2 are independently H, hydroxy or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted by 1 to 4 substituents each independently selected from the group consisting of hydroxyl, COOH and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1 to 4 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, in the compound of formula (I-3), one of ^RA1 and ^RA2 is H and the other of ^RA1 and ^RA2 is hydroxy or optionally substituted (C ₁ -C ₆ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₆ alkyl)oxy-group is optionally substituted by 1 to 4 substituents each independently selected from the following: hydroxyl, COOH and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1 to 4 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, in the compound of formula (I-3), one of ^RA1 and ^RA2 is H and the other of ^RA1 and ^RA2 is hydroxy or optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted by 1 to 2 substituents independently selected from the following: hydroxyl and optionally substituted phenyl,

The optionally substituted phenyl group is optionally substituted by 1-2 substituents each independently selected from the following substituents: (C ₁ -C ₄ alkyl)oxy-.

In one embodiment, in the compound of formula (I-3), ^RA1 and ^RA2 are each independently optionally substituted ( _C1 - _C4 alkyl)oxy-, wherein the alkyl group of the optionally substituted ( _C1 - _C4 alkyl)oxy- is optionally substituted with 1-2 hydroxy substituents.

In one embodiment, in the compound of formula (I-3), R ³ and R ⁵ are -CO-NH ₂ .

In one embodiment, in the compound of formula (I-3), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

The substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: halogen and C _1-4 alkyl.

In one embodiment of the compound of formula (I-3), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

The substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: fluorine and C _1-2 alkyl.

In one embodiment of the compound of formula (I-3), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 fluorine substituents.

In one embodiment of the compound of formula (I-3), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

The substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents each independently being a C _1-2 alkyl group.

In one embodiment of the compound of formula (I-3), B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 hydroxy substituents.

In one embodiment of the compounds of formula (I-3), B is -CH ₂ -CH ₂ - substituted with 1-2 hydroxy substituents.

In one embodiment, the compound of formula (I-3), wherein ^Rx and ^Ry are each independently methyl or ethyl. In another embodiment, the compound of formula (I-3), wherein both ^Rx and ^Ry are methyl. In another embodiment, the compound of formula (I-3), wherein one of ^Rx and ^Ry is methyl and the other is H.

In one embodiment, the compound of formula (I-3), wherein

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl;

One of R ^A1 and R ^A2 is H and the other of R ^A1 and R ^A2 is optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted by 1 to 2 substituents each independently selected from the following: hydroxyl and optionally substituted phenyl,

wherein the optionally substituted phenyl is optionally substituted by 1-2 substituents each independently selected from the following: (C ₁ -C ₄ alkyl)oxy-;

^R3 and ^R5 are both -CO- _NH2 ; and

B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: fluorine and C _1-2 alkyl; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

Or both ^RX and ^Ry are independently _C1 - _C4 alkyl.

In one embodiment, the compound of formula (I-3), wherein

R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl;

One of R ^A1 and R ^A2 is H and the other of R ^A1 and R ^A2 is optionally substituted (C ₁ -C ₄ alkyl)oxy-,

wherein the alkyl group of the optionally substituted (C ₁ -C ₄ alkyl)oxy-group is optionally substituted with 1-2 hydroxy substituents;

^R3 and ^R5 are both -CO- _NH2 ; and

B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-,

wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1 to 4 substituents independently selected from the following: hydroxyl and C _1-2 alkyl; and

At least one of ^RX or ^Ry is independently C ₁ -C ₄ alkyl and the other is H,

Or both ^RX and ^Ry are independently _C1 - _C4 alkyl.

Representative compounds of the present invention include compounds of the embodiments. It should be understood that the present invention encompasses compounds of formula (I) as free bases and as salts thereof (e.g., as pharmaceutically acceptable salts thereof). In one embodiment, the present invention relates to compounds of formula (I) in the form of free bases. In another embodiment, the present invention relates to compounds of formula (I) in the form of salts, particularly pharmaceutically acceptable salts. It should be further understood that, in one embodiment, the present invention relates to compounds of the embodiments in the form of free bases. In another embodiment, the present invention relates to compounds of the embodiments in the form of salts, particularly pharmaceutically acceptable salts.

Specific embodiments of the compounds of the present invention include:

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(2E,2'E)-1,1'-(pentane-1,5-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-methoxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((E)-4-((E)-7-(benzyloxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((E)-4-((E)-4-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-(4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-(((4R,5R)-5-(((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-(4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

4-(((E)-6-carbamoyl-3-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-methyl-2,3-dihydro-1H-benzo[d]imidazol-4-yl)oxy)butanoic acid methyl ester,

(E)-1-((E)-4-((E)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-7-(3-(dimethylamino)propoxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((3-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxylic acid methyl ester,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-7-bromo-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(2E,2'E)-1,1'-(2,3-dihydroxybutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide),

(2E,2'E)-1,1'-((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide),

(E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(2E,2'E)-1,1'-((2R,3R)-2,3-diethoxybutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide),

(E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-phenethyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-7-bromo-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-(5-((E)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pentyl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pentyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-(5-((E)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)pentyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide,

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxylic acid butyl ester

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxylic acid,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-4-(morpholinomethyl)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((2-fluorobenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-fluorobenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((3-fluorobenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-7-isobutoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-7-(3-(dimethylamino)propoxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-1-((E)-4-((E)-7-(3-(dimethylamino)propoxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-7-(2-hydroxy-2-methylpropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(2E,2'E)-1,1'-((meso)-2,3-dimethoxybutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide),

(E)-1-((E)-4-((E)-7-(3-(4,4-difluoropiperidin-1-yl)propoxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide,

(5aE,21E,29E)-8-ethyl-26-(3-hydroxypropoxy)-5,10,18,22,29,30-hexamethyl-7,20-dioxo-5,7,8,11,12,13,14,15,20,22,28,31-dodecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-l][1,3,6,15,17]pentaazacyclohenicosine-3,24-dicarboxamide,

or a tautomer thereof;

or a salt thereof, in particular a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of the above formula, such as the compound of formula (I), formula (I-2) or formula (I-3) is (E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide having the following structure

or a tautomer thereof;

or a salt thereof, in particular a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of the above formula, such as the compound of formula (I), formula (I-2) or formula (I-3) is (E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide having the following structure

or a tautomer thereof;

or a salt thereof, in particular a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of the above formula, such as the compound of formula (I), formula (I-2) or formula (I-3), is not the following compound: (E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide having the following structure

or a tautomer thereof;

or a salt thereof, in particular a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of the above formula, such as the compound of formula (I), formula (I-2) or formula (I-3), is not the following compound: (E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide having the following structure

or a tautomer thereof;

or a salt thereof, in particular a pharmaceutically acceptable salt thereof.

The compounds of the invention may contain one or more asymmetric centers (also referred to as chiral centers), such as chiral carbon or chiral -SO- moieties. The compounds of the invention containing one or more chiral centers may exist as racemic mixtures, diastereomeric mixtures, enantiomerically enriched mixtures, diastereoisomerically enriched mixtures, or as enantiomerically pure or diastereoisomerically pure individual stereoisomers.

The stereochemistry of the chiral centers present in the compounds of the present invention is generally indicated in the compound names and/or chemical structures illustrated herein. Where the stereochemistry of the chiral centers present in the compounds of the present invention or in any chemical structure illustrated herein is not specified, the structure is intended to encompass any stereoisomers and all mixtures thereof. Thus, the present invention encompasses all isomers of compounds of formula (I) and their salts, whether as individual isomers separated so as to be substantially free of other isomers (i.e., pure) or as mixtures (i.e., racemates and racemic mixtures). Individual isomers separated so as to be substantially free of other isomers (i.e., pure) may be separated so that there is less than 10%, particularly less than about 1%, for example less than about 0.1% of other isomers.

Individual stereoisomers of the compounds of the invention (or mixtures that may be enriched in stereoisomers) may be resolved using methods known to those skilled in the art. For example, such resolution may be performed as follows: (1) by forming diastereomeric salts, complexes, or other derivatives; (2) by selective reaction with stereoisomer-specific reagents, such as by enzymatic oxidation or reduction; or (3) by gas-liquid chromatography or liquid chromatography in a chiral environment, such as in the presence of a chiral support (such as silica with bound chiral ligands) or in a chiral solvent. It should be understood that in the case where the desired stereoisomer is converted into another chemical entity by one of the above separation methods, another step is required to release the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents, or by converting one enantiomer into another enantiomer by asymmetric transformation.

The present invention also includes various deuterated forms of the compounds of the present invention. Each available hydrogen atom bound to a carbon atom can be independently replaced by a deuterium atom. One of ordinary skill in the art will know how to synthesize deuterated forms of the compounds of the present invention. For example, α-deuterated α-amino acids are commercially available or can be prepared by conventional techniques ( see, for example: Elemes, Y. and Ragnarsson, UJ Chem. Soc., Perkin Trans. 1, 1996, 6, 537-40 ). Using such compounds can allow the preparation of compounds in which the hydrogen atom at the chiral center is replaced by a deuterium atom. Other commercially available deuterated starting materials can be used to prepare deuterated analogs of compounds of the invention (see, for example, methyl- d3 _- amine available from Aldrich Chemical Co., Milwaukee, WI), or they can be synthesized using conventional techniques employing deuterated reagents (e.g., by reduction using lithium aluminum deuteride or sodium borodeuteride or by metal-halogen exchange followed by quenching with _D2O or _methanol - d3 ).

Suitable pharmaceutically acceptable salts of compounds of formula (I) may include acid addition salts or base addition salts. For a review of suitable pharmaceutically acceptable salts, see Berge et al. , J. Pharm. Sci., 66: 1-19, (1977) and PH Stahl and CG Wermuth, eds., Handbook of Pharmaceutical Salts: Properties, Selection and Use , Weinheim/Zürich: Wiley-VCH/VHCA (2002).

Salts of compounds of formula (I) containing basic amines or other basic functional groups may be prepared by any suitable method known in the art, such as treating the free base with a suitable inorganic or organic acid. Examples of pharmaceutically acceptable salts so formed include acetates, adipates, ascorbates, aspartates, benzenesulfonates, benzoates, camphorates, camphor-sulfonates (camsylate), caprates (decanoate), caproates (hexanoate), carprylates (octanoate), carbonates, bicarbonates, cinnamates, citrates, cyclohexylsulfamates, dodecyl sulfates (estolate), ethane -1,2-disulfonate (edisylate), ethanesulfonate (esylate), formate, fumarate (hemifumarate, etc.), galactonate (mucate), gentisate (2,5-dihydroxybenzoate), glucoheptonate (gluceptate), gluconate, glucuronate, glutamate, glutarate, glycerophosphate, glycolate, hippurate, hydrobromide, hydrochloride (dihydrochloride, etc.), hydroiodide, isobutyrate, lactate, lactobionate, laurate, maleate, malate, malonate, mandelate, methanesulfonate (mesylat e)), naphthalene-1,5-disulfonate (naphthalenedisulfonate), naphthalene-sulfonate (napsylate), nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, phosphate (diphosphate, etc.), propionate, pyroglutamate, salicylate, sebacate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate (tosylate), undecylenate, 1-hydroxy-2-naphthoate, 2,2-dichloroacetate, 2-hydroxyethanesulfonate (isethionate), 2-oxoglutarate, 4-acetamidobenzoate and 4-aminosalicylate.

Salts of disclosed compounds containing carboxylic acids or other acidic functional groups can be prepared by reaction with a suitable base. Such pharmaceutically acceptable salts can be prepared with bases that provide pharmaceutically acceptable cations, including alkali metal salts (particularly sodium and potassium salts), alkaline earth metal salts (particularly calcium and magnesium salts), aluminum salts and ammonium salts, and salts prepared from physiologically acceptable organic bases, such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N , N' -dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tris-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, N , N' -bisdehydroabietylamine, glucosamine, N -methylglucosamine, colidine, choline, quinine, quinoline and basic amino acids, such as lysine and arginine.

The present invention includes within its scope all possible stoichiometric and non-stoichiometric forms of the salts (eg, hydrobromide, dihydrobromide, fumarate, hemifumarate, etc.) of the compounds of formula (I).

When a disclosed compound or salt thereof is named or depicted by a structure, it is understood that the compound or salt, including solvates thereof (particularly hydrates), may exist in a crystalline form, a non-crystalline form, or a mixture thereof. The compound or its salt or solvate (particularly hydrate) may also exhibit polymorphism (i.e., the ability to exist in different crystalline forms). These different crystalline forms are generally referred to as "polymorphs". It is understood that the present invention includes all polymorphs of any compound of the present invention, such as all polymorphic forms of any compound named or depicted by a structure herein, including any salt and/or solvate (particularly hydrate) thereof.

Polymorphs have the same chemical composition, but the characteristics of packing, geometric arrangement and other descriptive crystalline solids are different. Therefore, polymorphs can have different physical properties, such as shape, density, hardness, deformability, stability and solubility characteristics. Polymorphs typically show different melting points, IR spectra and X-ray powder diffraction patterns, which can be used for identification. It should be understood that different polymorphs can be produced, for example, by changing or adjusting the conditions used in the compound described in crystallization/recrystallization. Polymorphic forms can be characterized and distinguished using a variety of conventional analytical techniques, including but not limited to, X-ray powder diffraction (XRPD) spectrum, infrared (IR) spectrum, Raman spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and solid-state nuclear magnetic resonance (SSNMR).

The skilled person will appreciate that pharmaceutically acceptable solvates (particularly hydrates) of compounds of formula (I), including pharmaceutically acceptable solvates of pharmaceutically acceptable salts of compounds of formula (I), may be formed when solvent molecules are incorporated into the crystal lattice during crystallization. Solvates may involve non-aqueous solvents such as ethanol, or they may involve water as a solvent incorporated into the crystal lattice. Solvates in which water is a solvent incorporated into the crystal lattice are typically referred to as "hydrates".

The present invention includes within its scope all possible stoichiometric and non-stoichiometric salt and/or hydrate forms.

Salts and solvates (e.g., hydrates and hydrates of salts) of the compounds of the invention suitable for use in medicine are those in which the counterion or associated solvent is pharmaceutically acceptable. Salts with non-pharmaceutically acceptable counterions are also within the scope of the invention, for example, used as intermediates in the process of preparing other compounds of the invention.

In general, pharmaceutically acceptable salts can be readily prepared by using the desired acid or base as appropriate. The resulting salt may be crystallized or precipitated from solution, or formed by trituration, and may be recovered by filtration or by evaporation of the solvent.

Since the compounds of the present invention are intended for use in pharmaceutical compositions, it will be readily appreciated that each of them is preferably provided in substantially pure form, e.g., at least 60% pure, more suitably at least 75% pure, and preferably at least 85%, especially at least 98% pure (% on a weight-to-weight basis). Impure preparations of the compounds may be used to prepare purer forms for use in pharmaceutical compositions.

The present invention encompasses all prodrugs of the compounds of the invention which, upon administration to a recipient, are capable of providing (directly or indirectly) a compound of the invention or its active metabolite or residue. Such derivatives are identifiable to those skilled in the art without undue experimentation. Nevertheless, reference is made to the teachings of Burger's Medicinal Chemistry and Drug Discovery, 5th Edition, Vol. 1: Principles and Practice (which is incorporated herein by reference to the extent that such derivatives are taught).

It should be further understood that the present invention includes within its scope all tautomeric or isomeric forms of any free base form of the compounds of the present invention and all possible stoichiometric and non-stoichiometric salt forms. The compounds of the present invention can be used to treat or prevent diseases and conditions in which modulation of STING is beneficial. Such STING-mediated diseases and conditions include inflammation, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, metabolic diseases and cardiovascular diseases. The compounds of the present invention can also be used as immunogenic compositions or vaccine adjuvants. Therefore, the present invention relates to a method of modulating STING, which comprises contacting a cell with a compound of the present invention.

One aspect of the invention provides methods for treating or preventing STING-mediated diseases and conditions in which agonizing STING is beneficial. Exemplary diseases/conditions include, but are not limited to, cancer, infectious diseases (e.g., HIV, HBV, HCV, HPV, and influenza), vaccine adjuvants.

One aspect of the invention provides methods for treating or preventing a STING-mediated disease or condition in which inhibition of STING is beneficial. Exemplary diseases/conditions include, but are not limited to, systemic lupus erythematosus (SLE), cutaneous lupus, lupus nephritis, psoriasis, diabetes including insulin-dependent diabetes mellitus (IDDM), obesity-related insulin resistance and non-alcoholic fatty liver disease (NAFLD), dermatomyositis, systemic sclerosis (scleroderma) and Sjögren's syndrome (SS), rheumatoid arthritis, psoriatic arthritis, STING-associated vasculitis of infancy (SAVI), Aicardi Goutieres syndrome (AGS), pernio lupus, mixed connective tissue disease, neuroinflammation associated with Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, Huntington's disease and multiple sclerosis, and cardiac inflammation associated with myocardial infarction.

In one embodiment, the present invention provides a compound of the present invention for use in therapy. The present invention also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in therapy. The present invention particularly provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating a STING-mediated disease or condition.

The present invention also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is used as a vaccine adjuvant. Therefore, an immunogenic composition or vaccine adjuvant is also provided, which comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In a further embodiment of the present invention, there is provided a composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more immunostimulants.

In another embodiment, the present invention provides a compound of the present invention for use in treating a STING-mediated disease or condition and/or as an immunogenic composition or vaccine adjuvant. In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in improving organ damage or injury suffered due to a STING-mediated disease or condition.

The present invention further provides the use of the compounds of the present invention in the preparation of a medicament for treating a disease or condition mediated by STING. The present invention further provides the use of a compound of formula (I) or a salt thereof, in particular a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating a disease or condition mediated by STING, such as the diseases and conditions described herein.

The present invention further provides the use of a compound of formula (I) or a salt thereof, particularly a pharmaceutically acceptable salt thereof, in the preparation of a vaccine. Further provided is the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of an immunogenic composition comprising an antigen or an antigenic composition, the immunogenic composition being used to treat or prevent a disease. Further provided is the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a vaccine composition comprising an antigen or an antigenic composition, the vaccine composition being used to treat or prevent a disease.

In another embodiment, the present invention relates to a method for treating a STING-mediated disease or condition, comprising administering a therapeutically effective amount of a compound of the present invention to a person in need thereof. In another embodiment, the present invention relates to a method for treating a STING-mediated disease or condition, comprising administering a therapeutically effective amount of a compound of formula (I) or a salt thereof, particularly a pharmaceutically acceptable salt thereof, to a person in need thereof.

In another embodiment, the present invention relates to a method for treating or preventing a disease, comprising administering an immunogenic composition comprising an antigen or antigenic composition and a compound of formula (I) or a pharmaceutically acceptable salt thereof to a human subject suffering from or susceptible to the disease. In another embodiment, the present invention relates to a method for treating or preventing a disease, comprising administering a vaccine composition comprising an antigen or antigenic composition and a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient human subject suffering from or susceptible to the disease.

In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating inflammation. In a further aspect, a method for treating inflammation is provided, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a person in need thereof. In a further aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in preparing a medicament for treating inflammation.

In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating allergic diseases. In a further aspect, a method for treating allergic diseases is provided, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a person in need thereof. In a further aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in preparing a medicament for treating allergic diseases. Exemplary allergic diseases include allergic rhinitis, hay fever, atopic dermatitis, urticaria.

In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating an autoimmune disease. In a further aspect, a method for treating an autoimmune disease is provided, comprising administering to a person in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In a further aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in preparing a medicament for treating an autoimmune disease.

In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating infectious diseases. In a further aspect, a method for treating infectious diseases is provided, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a person in need thereof. In a further aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in preparing a medicament for treating an infectious disease.

In one embodiment, the invention relates to a method of treating HIV infection in a human by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the invention relates to a method of treating HIV infection in a human by administering to a human having HIV infection or at risk of having HIV infection a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In another embodiment, the invention relates to a method of treating AIDS infection in a human having AIDS infection by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In one embodiment, the invention relates to a method of treating HBV infection in a human by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the invention relates to a method of treating HBV infection in a human having HBV infection or at risk of having HBV infection by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In one embodiment, the method for treating HBV infection includes administering a first therapeutic agent. In one embodiment, the method includes administering a first therapeutic agent, wherein the first therapeutic agent is a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and administering one or more second therapeutic agents. In one embodiment, the first therapeutic agent and one or more second therapeutic agents are co-administered.

In one embodiment, the first therapeutic agent and one or more second therapeutic agents are co-administered sequentially or concomitantly. In one embodiment, one or more second therapeutic agents are also compounds of formula (I). In one embodiment, one or more second therapeutic agents are different from compounds or compositions described herein. Examples of one or more second therapeutic agents include, but are not limited to, anti-inflammatory agents, chemotherapeutics, or anti-infective agents. In other related embodiments, additional therapeutic agents may be HBV agents, HCV agents, chemotherapeutics, antibiotics, analgesics, non-steroidal anti-inflammatory (NSAID) agents, antifungal agents, antiparasitic agents, anti-nausea agents, antidiarrheal agents, immunomodulators, or immunosuppressants.

In one embodiment, the one or more second therapeutic agents are HBV agents. In one embodiment, HBV agents may include, but are not limited to, interferon α-2b, interferon 5 α-2a, and interferon αcon-1 (pegylated and unpegylated), ribavirin; HBV RNA replication inhibitors; HBV antigen production inhibitors; HBV therapeutic vaccines; HBV preventive vaccines; lamivudine (3TC); entecavir (ETV); tenofovir disoproxil fumarate (TDF); telbivudine (LdT); adefovir; or HBV antibody therapy (monoclonal or polyclonal).

In one embodiment, the one or more second therapeutic agents are HCV agents. In one embodiment, HCV agents may include, but are not limited to, interferon alpha-2b, interferon alpha-2a, and interferon alphacon-1 (pegylated and unpegylated); ribavirin; HCV RNA replication inhibitors (e.g., ViroPharma's VP50406 series); HCV antisense agents; HCV therapeutic vaccines; HCV protease inhibitors; HCV helicase inhibitors; or HCV monoclonal or polyclonal antibody therapies.

In one embodiment, one or more second therapeutic agents are anti-inflammatory agents (i.e., treatments that reduce inflammation). In one embodiment, treatments that reduce inflammation may include, but are not limited to, therapeutic lifestyle changes, steroids, NSAIDs, or DMARDs. The steroids delivered may be corticosteroids. The NSAIDs may be aspirin, acetaminophen, ibuprofen, naproxen, COX inhibitors, indomethacin, etc. The DMARDs may be TNF inhibitors, purine synthesis inhibitors, calcineurin inhibitors, pyrimidine synthesis inhibitors, sulfasalazine, methotrexate, etc.

In one embodiment, the one or more second therapeutic agents are chemotherapeutic agents (i.e., cancer therapeutic agents). Chemotherapeutic agents may include, but are not limited to, daunorubicin, daunomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide, ifosfamide, cytarabine, bis-chloroethylnitrosurea, busulfan, mitomycin C, actinomycin D, mithramycin, prednisone, hydroxyprogesterone, testosterone, tamoxifen, dacarbazine, procarbazine, hexamethylmelamine, pentamethylmelamine, mitoxantrone , amsacrine, chlorambucil, methylcyclohexylnitrosurea, nitrogen mustard, melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine (CA), 5-azacytidine, hydroxyurea, deoxycoformycin, 4-hydroxyperoxycyclophosphoramide, 5-fluorouracil (5-FU), 5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX), colchicine, paclitaxel, vincristine, vinblastine, etoposide, trimetrexate, teniposide, cisplatin, gemcitabine, and diethylstilbestrol (DES).

In one embodiment, one or more second therapeutic agents are immunomodulators, known as natural immune active agents, checkpoint inhibitors, T-cell stimulators or other agents that restore adaptive immune responses to HBV. Immunomodulators include, but are not limited to, antibodies or small molecules that antagonize CTLA-4, such as Ipilimumab (YERVOY), PD-1, such as Opdivo/nivolumab and Keytruda/pembrolizumab), PDL1 such as TECENTRIQ™ (atezumab), LAG3, TIM3 or IDO. Immunomodulators include, but are not limited to, antibodies or small molecules that stimulate ICOS, OX-40, TLRs, IL7R or IL12R.

In one embodiment, the one or more second therapeutic agents are anti-infective agents. Examples of anti-infective agents include, but are not limited to, antibiotics, antifungal drugs, and antiviral drugs.

In one embodiment, the invention relates to a method of treating HCV infection in a human by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the invention relates to a method of treating HCV infection in a human having HCV infection or at risk of having HCV infection by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In one embodiment, the invention relates to a method of treating influenza in a human by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the invention relates to a method of treating influenza in a human having an infection or at risk of having an infection by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In one embodiment, the invention relates to a method for treating human papillomavirus (HPV) infection in a human by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the invention relates to a method for treating HPV infection in a human having HPV infection or at risk of having HPV infection by administering to the human a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

As used herein, the terms "cancer", "neoplasm" and "tumor" are used interchangeably and, in the singular or plural, refer to cells that undergo malignant transformation, which makes them pathological to the host organism. Primary cancer cells can be easily distinguished from non-cancerous cells by well-established techniques, particularly histological examination. As used herein, the definition of cancer cells includes not only primary cancer cells, but also any cells derived from cancer cell ancestors. This includes metastatic cancer cells, as well as in vitro cultures and cell lines derived from cancer cells. When referring to a type of cancer that usually appears as a solid tumor, a "clinically detectable" tumor is a tumor that is detectable based on tumor mass; for example, by procedures such as computed tomography (CT) scans, magnetic resonance imaging (MRI), X-rays, ultrasound or palpation during a physical examination, and/or a tumor that is detectable due to the expression of one or more cancer-specific antigens in a sample that can be obtained from a patient. The tumor can be a hematopoietic (or blood or hematological or blood-related) cancer, for example, a cancer derived from blood cells or immune cells, which can be referred to as a "liquid tumor". Specific examples of hematological neoplasm-based clinical conditions include leukemias such as chronic myeloid leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, and acute lymphocytic leukemia; plasma cell malignancies such as multiple myeloma, MGUS, and Waldenstrom's macroglobulinemia; lymphomas such as non-Hodgkin's lymphoma, Hodgkin's lymphoma, and the like.

Cancer can be any cancer in which there is an abnormal number of blasts or unwanted cell proliferation or diagnosed as a blood cancer, including lymphoid and myeloid malignancies. Myeloid malignancies include, but are not limited to, acute myeloid (or myeloid cell or myeloid or myeloblastic) leukemia (undifferentiated or differentiated), acute promyelocytic (or promyelocytic or promyelogenous or promyeloblastic) leukemia, acute myelomonocytic (or myelomonocytic (myelomonoblastic)) leukemia, acute monocytic (or monoblastic (monoblastic)) leukemia, erythroleukemia, and megakaryoblastic (or megakaryoblastic) leukemia. These leukemias can be collectively referred to as acute myeloid (or myelocytic (myelocytic) or myeloid (myelogenous)) leukemia (AML). Myeloid malignancies also include myeloproliferative disorders (MPDs), including, but not limited to, chronic myeloid (or myeloid) leukemia (CML), chronic myelomonocytic leukemia (CMML), idiopathic thrombocythemia (or thrombocythemia), and polycythemia vera (PCV). Myeloid malignancies also include myelodysplasia (or myelodysplastic syndrome or MDS), which may be referred to as refractory anemia (RA), refractory anemia with excess blasts (RAEB), and refractory anemia with excess blasts in transformation (RAEBT); and myelofibrosis (MFS) with or without unexplained myeloid metaplasia.

Hematopoietic cancers also include lymphoid malignancies, which can affect the lymph nodes, spleen, bone marrow, peripheral blood, and/or extranodal sites. Lymphoid cancers include B-cell malignancies, which include, but are not limited to, B-cell non-Hodgkin's lymphoma (B-NHL). B-NHL can be indolent (or low grade), intermediate (or aggressive), or high grade (very aggressive). Indolent B-cell lymphomas include follicular lymphoma (FL); small lymphocytic lymphoma (SLL); marginal zone lymphoma (MZL), including lymph node MZL, extranodal MZL, splenic MZL, and splenic MZL with villous lymphocytes; lymphoplasmacytic lymphoma (LPL); and mucosa-associated lymphoid tissue (MALT or extranodal marginal zone) lymphoma. Intermediate-grade B-NHL includes mantle cell lymphoma (MCL) with or without leukemic involvement, diffuse large cell lymphoma (DLBCL), follicular large cell (or grade 3 or 3B) lymphoma, and primary mediastinal lymphoma (PML). High-grade B-NHL includes Burkitt's lymphoma (BL), Burkitt-like lymphoma, small non-cleaved cell lymphoma (SNCCL), and lymphoblastic lymphoma. Other B-NHLs include immunoblastic lymphoma (or immunocytoma), primary effusion lymphoma, HIV-related (or AIDS-related) lymphoma, and post-transplant lymphoproliferative disorder (PTLD) or lymphoma. B-cell malignancies also include, but are not limited to, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), Waldenstrom's macroglobulinemia (WM), hairy cell leukemia (HCL), large granular lymphocytic (LGL) leukemia, acute lymphoid (or lymphocytic or lymphoblastic) leukemia, and Castleman's disease. NHL can also include T-cell non-Hodgkin's lymphoma (T-NHL), which includes but is not limited to non-Hodgkin's lymphoma not otherwise specified (NOS), peripheral T-cell lymphoma (PTCL), anaplastic large cell lymphoma (ALCL), angioimmunoblastic lymphoid disorder (AILD), nasal natural killer (NK) cell/T-cell lymphoma, gamma/delta lymphoma, cutaneous T-cell lymphoma, mycosis fungoides, and Sezary syndrome.

Hematopoietic cancer also includes Hodgkin's lymphoma (or disease), including classical Hodgkin's lymphoma, nodular sclerosing Hodgkin's lymphoma, mixed cell type Hodgkin's lymphoma, lymphocyte dominant (LP) Hodgkin's lymphoma, nodular LP Hodgkin's lymphoma and lymphocyte depleted Hodgkin's lymphoma. Hematopoietic cancer also includes plasma cell diseases or cancers, such as multiple myeloma (MM), including smoldering MM, monoclonal gammopathy of undetermined significance (or unknown or unclear) (MGUS), plasmacytoma (bone, extramedullary), lymphoplasmacytic lymphoma (LPL), Waldenstrom's macroglobulinemia, plasma cell leukemia and primary amyloidosis (AL). Hematopoietic cancers may also include other cancers of additional hematopoietic cells, including polymorphonuclear leukocytes (or neutrophils), basophils, eosinophils, dendritic cells, platelets, erythrocytes, and natural killer cells. Tissues including hematopoietic cells referred to herein as "hematopoietic tissues" include bone marrow; peripheral blood; thymus; and peripheral lymphoid tissues, such as spleen, lymph nodes, lymphoid tissue associated with mucosa (such as gut-associated lymphoid tissue), tonsils, Peyer's patches, and appendix, as well as lymphoid tissue associated with other mucosa (e.g., bronchial lining).

In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating cancer and precancerous syndromes. In a further aspect, a method for treating cancer and precancerous syndromes is provided, comprising administering to a person in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In a further aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in preparing a medicament for treating cancer and precancerous syndromes.

Related autoimmune diseases include, but are not limited to, STING-associated vasculitis of infancy (SAVI), Aicardi Goutieres syndrome (AGS), pernio lupus, ataxia telangiectasia (also known as Louis-Bar syndrome), retinal vasculopathy with cerebral leukodystrophy (RCVL), systemic lupus erythematosus (SLE), cutaneous lupus, lupus nephritis, psoriasis, diabetes (including insulin-dependent diabetes mellitus (IDDM)), dermatomyositis, human immunodeficiency virus (HIV), AIDS, polymyositis, systemic sclerosis (scleroderma) and Sjögren's syndrome (SS), rheumatoid arthritis Arthritis, psoriatic arthritis, polyarthritis, osteoarthritis, myasthenia gravis, polyarteritis nodosa, vasculitis, cutaneous vasculitis, antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, Henoch-Schönlein purpura, autoimmune hepatitis, primary sclerosing cholangitis, Wegener's granulomatosis, microscopic polyangiitis, Behcet's disease, spondylitis, giant cell arteritis, polymyalgia rheumatica, Raynaud's phenomenon, primary biliary cirrhosis, primary vasculitis of the central nervous system, microscopic polyangiitis, neuromyelitis optica, and mixed connective tissue disease.

Inflammation represents a group of vascular, cellular and neural responses to trauma. Inflammation can be characterized by the movement of inflammatory cells such as monocytes, neutrophils and granulocytes into tissues. This is usually associated with reduced endothelial barrier function and edema into tissues. Inflammation can be divided into acute or chronic. Acute inflammation is the body's initial response to harmful stimuli and is achieved by increased movement of plasma and leukocytes from the blood to damaged tissues. A cascade of biochemical events propagates the inflammatory response and matures it, which involves the local vascular system, immune system and various cells in the damaged tissue. Persistent inflammation, known as chronic inflammation, causes progressive changes in the cell types present at the site of inflammation and is characterized by simultaneous destruction and healing of tissues from the inflammatory process.

Inflammation can be beneficial and is usually self-limiting when it occurs as part of an immune response to infection or as an acute response to trauma. However, inflammation can be harmful under a variety of conditions. This includes excessive inflammation in response to infectious agents, which can lead to significant organ damage and death (e.g., in the setting of sepsis). In addition, chronic inflammation is usually harmful and is the root cause of many chronic diseases, causing severe and irreversible damage to tissues. In such cases, the immune response is usually directed against one's own tissues (autoimmunity), although chronic responses to foreign entities may also result in bystander damage to one's own tissues.

Therefore, the goal of anti-inflammatory therapy is to reduce this inflammation to suppress autoimmunity when present and allow physiological processes or healing and tissue repair to proceed.

The compounds of the invention can be used to treat inflammation of any tissue or organ of the body, including musculoskeletal inflammation, vascular inflammation, neurological inflammation, digestive system inflammation, ocular inflammation, cardiac inflammation, adipose tissue inflammation, reproductive system inflammation, and other inflammations, as exemplified below.

Musculoskeletal inflammation refers to any inflammatory condition of the musculoskeletal system, particularly those affecting skeletal joints, including those of the hands, wrists, elbows, shoulders, jaws, spine, neck, hips, knees, ankles and feet, and conditions affecting tissues connecting muscle to bone, such as tendons. Examples of musculoskeletal inflammation that can be treated with the compounds of the invention include arthritis (including, for example, osteoarthritis, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, acute and chronic infectious arthritis, arthritis associated with gout and pseudogout, and juvenile idiopathic arthritis), tendinitis, synovitis, tenosynovitis, bursitis, fibrositis (fibromyalgia), epicondylitis, myositis, and osteitis (including, for example, Paget's disease, osteitis pubis, and osteitis fibrosa cystica).

Ocular inflammation refers to inflammation of any structure of the eye, including the eyelids. Examples of ocular inflammation that can be treated with the compounds of the invention include blepharitis, blepharoptosis, conjunctivitis, dacryoadenitis, keratitis, keratoconjunctivitis sicca (dry eyes), scleritis, trichiasis, and uveitis.

Examples of inflammatory diseases of the nervous system that may be treated with the compounds of the invention include encephalitis, Guillain-Barré syndrome, meningitis, neuromyotonia, narcolepsy, multiple sclerosis, myelitis, CNS vasculitis, and schizophrenia.

Examples of inflammation associated with neurodegenerative diseases that may be treated with the compounds of the invention include Alzheimer's disease and related dementias, amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTD), Parkinson's disease and Huntington's disease.

Examples of inflammation of the vasculature or lymphatic system that may be treated with the compounds of the invention include arthrosclerosis, arthritis, phlebitis, vasculitis and lymphangitis.

Examples of inflammation of the cardiovascular system that may be treated with the compounds of the invention include, but are not limited to, myocardial infarction, heart failure, congenital heart defects, coronary artery disease, hypertension, cardiomyopathy, and other related cardiovascular conditions.

Examples of liver inflammation that can be treated with the compounds of the invention include, but are not limited to, liver fibrosis, alcoholic liver disease (ALD), non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis, and biliary liver disease.

Examples of adipose tissue inflammation that may be treated using the compounds of the invention include, but are not limited to, obesity and obesity-induced insulin resistance.

Examples of liver inflammation that may be treated with the compounds of the invention include, but are not limited to, liver fibrosis and fibromas, alcoholic liver disease (ALD), nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), and biliary liver disease.

Examples of pancreatic inflammation that may be treated using the compounds of the invention include, but are not limited to, pancreatitis and metabolic syndrome-induced pancreatic beta cell dysfunction.

Examples of renal inflammation that may be treated using the compounds of the invention include, but are not limited to, renal nephritis.

Examples of inflammatory diseases of the lung that may be treated using the compounds of the invention include pulmonary fibrosis, COPD and asthma.

Examples of inflammatory conditions of the eye that may be treated using the compounds of the invention include dry eye syndrome and age-related macular degeneration.

Examples of inflammatory disorders of the digestive system that may be treated with the compounds of the invention include cholangitis, cholecystitis, enteritis, enterocolitis, gastritis, gastroenteritis, inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), ileitis, and proctitis.

Examples of inflammatory disorders of the reproductive system that may be treated with the compounds of the invention include cervicitis, chorioamnionitis, endometritis, epididymitis, omphalitis, oophoritis, orchitis, salpingitis, tubo-ovarian abscess, urethritis, vaginitis, vulvitis, and vulvodynia.

In one embodiment, the compounds of the invention may be used to treat systemic lupus erythematosus, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease (IBD) (eg, Crohn's disease, ulcerative colitis).

The compounds of the invention are useful for treating autoimmune disorders with an inflammatory component. Such disorders include acute disseminated alopecia universalise, Behcet's disease, Chagas disease, STING-associated vasculitis of infancy (SAVI), Aicardi Goutieres syndrome (AGS), lupus pernio, ataxia telangiectasia (also known as Louis-Bar syndrome), retinal vasculopathy with cerebral leukodystrophy (RCVL), ANCA-associated vasculitis, chronic fatigue syndrome, dysautonomia, encephalomyelitis, ankylosing spondylitis, aplastic anemia, hidradenitis suppurativa, autoimmune hepatitis, autoimmune oophoritis, celiac disease, Crohn's disease, type 1 diabetes mellitus, giant cell arteritis, Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, Henoch-Schonlein purpura, Kawasaki disease, lupus erythematosus, microscopic colitis, microscopic polyarteritis, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, opsoclonus myoclonus syndrome syndrome, optic neuritis, Ord's thyroiditis, pemphigus, polyarteritis nodosa, polymyalgia, rheumatoid arthritis, Reiter's syndrome, Sjögren's syndrome, temporal arteritis, Wegener's granulomatosis, warm autoimmune hemolytic anemia, interstitial cystitis, Lyme disease, morphea, psoriasis, sarcoidosis, scleroderma, ulcerative colitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, and vitiligo.

The compounds of the invention are useful for treating T cell mediated hypersensitivity diseases with an inflammatory component. Such conditions include contact hypersensitivity, contact dermatitis (including that caused by poison ivy), urticaria, skin allergies, respiratory allergies (hay fever, allergic rhinitis) and gluten sensitive enteropathy (celiac disease).

Other inflammatory conditions that may be treated with the compounds of the invention include, for example, appendicitis, dermatitis, dermatomyositis, endocarditis, fibrositis, gingivitis, glossitis, hepatitis, hidradenitis suppurativa, iritis, laryngitis, mastitis, myocarditis, nephritis, otitis, pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis, pleurisy, pneumonia, prostatitis, pyelonephritis, and stomatitis, transplant rejection (involving organs such as the kidney, liver, heart, lung, pancreas (e.g., islet cells), bone marrow, cornea, small intestine, skin allografts, skin allografts, and heart valve xenografts, serum sickness, and graft-versus-host disease), acute pancreatitis, chronic pancreatitis, acute respiratory distress syndrome, Sezary syndrome, congenital adrenal hyperplasia, non-suppurative thyroiditis, hypercalcemia associated with cancer, pemphigus, bullous dermatitis herpetiformis, and stomatitis. dermatitisherpetiformis), severe erythema multiforme, exfoliative dermatitis, seborrheic dermatitis, seasonal or perennial allergic rhinitis, bronchial asthma, contact dermatitis, atopic dermatitis, drug hypersensitivity, allergic conjunctivitis, keratitis, herpes zoster ophthalmicus, iritis and iridocyclitis, chorioretinitis, optic neuritis, symptomatic sarcoidosis, fulminant or disseminated pulmonary tuberculosis chemotherapy, idiopathic thrombocytopenic purpura in adults, secondary thrombocytopenia in adults, acquired (autoimmune) hemolytic anemia, adult leukemia and lymphoma, acute leukemia in children, regional enteritis, autoimmune vasculitis, multiple sclerosis, chronic obstructive pulmonary disease, solid organ transplant rejection, sepsis. Preferred treatments include treatment of transplant rejection, rheumatoid arthritis, psoriatic arthritis, multiple sclerosis, type 1 diabetes, asthma, inflammatory bowel disease, systemic lupus erythematosus, psoriasis, chronic lung disease and inflammation associated with infectious conditions (e.g., sepsis). In one embodiment, the compounds of the invention are useful in the treatment of asthma.

Examples of cancer diseases and conditions in which the compounds of the invention may have a potentially beneficial anti-tumor effect include, but are not limited to, cancer of the lung, bone, pancreas, skin, head, neck, uterus, ovary, stomach, colon, breast, esophagus, small intestine, intestine, endocrine system, thyroid, parathyroid, adrenal glands, urethra, prostate, penis, testicles, ureter, bladder, kidney or liver; rectal cancer; anal region cancer; fallopian tube, endometrial, cervical, vagina, vulva, renal pelvis, renal cell; soft tissue sarcoma; myxoma; rhabdomyomas; fibromas; lipomas; teratomas; bile duct epithelial carcinomas; hepatoblastomas; angiosarcomas; hemangiomas; hepatomas; fibrosarcomas; chondrosarcomas; myeloma; chronic or acute leukemias; lymphocytic lymphomas; primary CNS lymphomas; tumors of the CNS; spinal cord (spinal cord) axis tumor; squamous cell carcinoma; synovial sarcoma; malignant pleural mesothelioma; brain stem glioma; pituitary adenoma; bronchial adenoma; enchondromatous hamartoma; mesothelioma; Hodgkin's disease, or a combination of one or more of the foregoing cancers.

Suitably, the present invention relates to a method for treating or lessening the severity of a cancer selected from the group consisting of brain cancer (glioma), glioblastoma, astrocytoma, glioblastoma multiforme, Bannayan-Zonana syndrome, Cowden disease, Lhermitte-Duclos disease, Wilman tumor, Ewing's sarcoma, rhabdomyosarcoma, ependymoma, medulloblastoma, head and neck cancer, kidney cancer, liver cancer, melanoma, ovarian cancer, pancreatic cancer, adenocarcinoma, ductal adenocarcinoma, adenosquamous carcinoma, acinar cell carcinoma, glucagonoma, insulinoma, prostate cancer, sarcoma, osteosarcoma, giant cell tumor of bone, thyroid cancer, lymphoblastic T-cell leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, hairy cell leukemia, acute myeloid leukemia, Lymphocytic leukemia, acute myeloid leukemia, chronic neutrophilic leukemia, acute lymphoblastic T-cell leukemia, plasmacytoma, immunoblastic large cell leukemia, mantle cell leukemia, multiple myeloma, megakaryoblastic leukemia, multiple myeloma, acute megakaryocytic leukemia, promyelocytic leukemia, erythroleukemia, malignant lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, lymphoblastic T-cell lymphoma, Burkitt's lymphoma, follicular lymphoma, neuroblastoma, bladder cancer, urothelial carcinoma, vulvar cancer, cervical cancer, endometrial cancer, renal cancer, mesothelioma, esophageal cancer, salivary gland cancer, hepatocellular carcinoma, gastric cancer, nasopharyngeal cancer, cheek cancer, oral cancer, GIST (gastrointestinal stromal tumor) and testicular cancer. In some embodiments, the compounds of the present invention can be used to treat solid or liquid tumors. In some embodiments, the compounds of the invention are useful for treating sarcomas, breast cancer, colorectal cancer, gastroesophageal cancer, melanoma, non-small cell lung cancer (NSCLC), clear cell renal cell carcinoma (RCC), lymphoma, squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma (HCC) and/or non-Hodgkin's lymphoma (NHL). Suitably, the present invention relates to a method of treating or lessening the severity of a precancerous syndrome in a mammal, including a human, wherein the precancerous syndrome is selected from the group consisting of cervical intraepithelial neoplasia, monoclonal gammopathy of undetermined significance (MGUS), myelodysplastic syndrome, aplastic anemia, cervical lesions, skin nevus (premelanoma), prostatic intraepithelial (intraductal) neoplasia (PIN), ductal carcinoma in situ (DCIS), colon polyps, and severe hepatitis or cirrhosis.

In one aspect, people have solid tumors. In one aspect, tumors are selected from head and neck cancer, gastric cancer, melanoma, renal cell carcinoma (RCC), esophageal cancer, non-small cell lung cancer, prostate cancer, colorectal cancer, ovarian cancer and pancreatic cancer. In one aspect, people have one or more of the following: colorectal cancer (CRC), esophageal cancer, cervical cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, melanoma, renal cell carcinoma (RCC), EC squamous cells, non-small cell lung cancer, mesothelioma and prostate cancer. In another aspect, people have liquid tumors, such as diffuse large B cell lymphoma (DLBCL), multiple myeloma, chronic lymphoblastic leukemia (CLL), follicular lymphoma, acute myeloid leukemia and chronic myeloid leukemia.

In one embodiment, the compounds of the invention are useful for treating skin cancer (e.g., non-melanoma skin cancer, squamous cell carcinoma, basal cell carcinoma) or actinic keratosis. In addition to the field effect for clearing superficial skin cancers, the compounds of the invention may also prevent the development of subsequent skin cancers and premalignant actinic keratoses in the treated patient.

The compounds of the invention may also be used to treat one or more diseases afflicting mammals characterized by cell proliferation in the field of disorders associated with neovascularization and/or vascular permeability, including vascular proliferative disorders, including arthritis (rheumatoid arthritis) and restenosis; fibrotic disorders, including cirrhosis and atherosclerosis; mesangial cell proliferative disorders, including glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndrome, proliferative retinopathy, organ transplant rejection and glomerulopathy; and metabolic disorders, including psoriasis, diabetes, chronic wound healing, inflammation and neurodegenerative diseases.

The compounds of the invention can be used to treat neurodegenerative diseases. Exemplary neurodegenerative diseases include, but are not limited to, multiple sclerosis, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and frontotemporal lobar degeneration (FTD).

The compounds of the present invention can be used to treat or prevent metabolic diseases (such as insulin resistance, non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH), obesity, diabetes, hypertension, fatty liver and cardiovascular disease.

The compounds of the present invention can be used to treat infectious diseases, which are any diseases caused by pathogen infection or occurring simultaneously with pathogen infection. Pathogens are broadly defined as any biological species that are foreign to the human tissue environment. Common pathogens that cause diseases include bacteria (many, such as TB), viruses (many, such as HBV, HIV, influenza viruses) and parasitic protozoa (such as Plasmodium falciparum that causes malaria). The compounds of the present invention can be used to treat infectious diseases derived from bacteria, such as TB infection (Mycobacterium tuberculosis), chlamydia infection, tularemia infection (Francisco tularemia), Plasmodium infection or infection from DNA or RNA viruses. The compounds of the present invention can be used to treat infectious diseases derived from the following DNA virus families: Herpesviridae (herpes simplex virus-1, Kaposi's sarcoma-associated virus and Epstein-Barr virus), Papillomaviridae (human papillomavirus), adenovirus and Hepadnaviridae (hepatitis B virus). Examples of RNA virus families include Retroviridae (human immunodeficiency virus), Flaviviridae (dengue virus, hepatitis C virus), Orthomyxoviridae (influenza), and Coronaviridae (human coronavirus and SARS coronavirus).

The compounds of the present invention can be used alone or in combination with other therapeutic agents. As modulators of immune responses, the compounds of the present invention can also be used in monotherapy or in combination with another therapeutic agent for the treatment of diseases and conditions in which the regulation of STING is beneficial. Therefore, the combination therapy according to the present invention comprises administering a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one other therapeutically active agent. In one embodiment, the combination therapy according to the present invention comprises administering at least one compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one other therapeutic agent. The compound of formula (I) and its pharmaceutically acceptable salt and the other therapeutic agent can be administered together or separately in a single pharmaceutical composition, and when administered separately, this can be performed simultaneously or sequentially in any order. The amount of the compound of formula (I) and its pharmaceutically acceptable salt and the other therapeutic agent and the relative timing of administration are selected to achieve the desired combined therapeutic effect. Therefore, in a further aspect, a combination product is provided, which comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more other therapeutic agents.

The compounds of formula (I) and their pharmaceutically acceptable salts can be used in combination with one or more other therapeutic agents useful for preventing or treating allergic diseases, inflammatory diseases or autoimmune diseases, such as: antigen immunotherapy, antihistamines, steroids, NSAIDs, bronchodilators (e.g., β2 agonists, adrenergic agonists, anticholinergics, theophylline), methotrexate, leukotriene modifiers and similar agents; monoclonal antibody therapy such as anti-IgE, anti-TNF, anti-IL-5, anti-IL-6, anti-IL-12, anti-IL-1 and similar agents; receptor therapy such as etanercept and similar agents; antigen nonspecific immunotherapy (e.g., interferon or other cytokines/chemokines, cytokine/chemokine receptor modulators, cytokine agonists or antagonists, TLR agonists and similar agents).

Formula (I) compound and its pharmaceutically acceptable salt can be used in combination with radiotherapy and/or surgery and/or at least one other therapeutic agent that can be used to treat cancer and precancerous syndrome. Any antitumor agent that is active for the susceptible tumor to be treated can be used in combination. Available typical antitumor agents include but are not limited to anti-microtubule agents, such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents, such as nitrogen mustards, oxazaphosphorines, alkyl sulfonates, nitrosoureas and triazenes; antibiotic agents, such as anthracyclines, actinomycins and bleomycins; topoisomerase II inhibitors, such as epipodophyllotoxins; antimetabolites, such as purine and pyrimidine analogs and antifolate compounds; topoisomerase I inhibitors, such as camptothecins; hormones and hormone analogs; signal transduction pathway inhibitors; non-receptor tyrosine angiogenesis inhibitors; immunotherapeutics; pro-apoptotic agents; cell cycle signal transduction inhibitors; immuno-oncology agents and immunostimulants.

Anti-microtubule or anti-mitotic agents are phase-specific agents that are active against the microtubules of tumor cells during the M phase or mitosis of the cell cycle. Examples of anti-microtubule agents include, but are not limited to, diterpenoids and vinca alkaloids.

Diterpenoids derived from natural sources are phase-specific anticancer agents that act during the _G2 /M phase of the cell cycle. It is believed that diterpenoids stabilize the β-tubulin subunit of microtubules by binding to this protein. The breakdown of the protein then appears to be inhibited, with a simultaneous arrest of mitosis followed by cell death. Examples of diterpenoids include, but are not limited to, paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytaxane-11-ene-9-one 4,10-diacetate 2-benzoate-13-[(2R,3S)-N-benzoyl-3-phenylisoserine ester], is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injection solution TAXOL®. It is a member of the taxane family of terpenes. Paclitaxel has been approved for clinical use in the United States for the treatment of refractory ovarian cancer (Markman et al., Yale Journal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann. Intem, Med., 111:273, 1989) and for the treatment of breast cancer (Holmes et al., J. Nat. Cancer Inst., 83:1797, 1991). It is a potential candidate for the treatment of skin tumors (Einzig et al., Proc. Am. Soc. Clin. Oncol., 20:46) and head and neck cancer (Forastire et al., Sem. Oncol., 20:56, 1990). The compound also shows potential for the treatment of polycystic kidney disease (Woo et al., Nature, 368:750. 1994), lung cancer and malaria. Treatment of patients with paclitaxel results in bone marrow suppression (multiple cell lineages, Ignoff, RJ et al., Cancer Chemotherapy Pocket Guide , 1998) associated with the duration of administration above the threshold concentration (50 nM) (Kearns, CM et al., Seminars in Oncology, 3 (6) p.16-23, 1995).

Docetaxel, (2R,3S)-N-carboxy-3-phenylisoserine, N-tert-butyl ester, 13-[5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytaxane-11-en-9-one 4-acetate 2-benzoate] trihydrate; commercially available as TAXOTERE® as an injection. Docetaxel is indicated for the treatment of breast cancer. Docetaxel is a semisynthetic derivative of paclitaxel qv prepared using the natural precursor 10-deacetyl-baccatin III extracted from the needles of the European yew tree.

Vinca alkaloids are stage-specific antitumor agents derived from plants of the genus Vinca. Vinca alkaloids act on the M phase (mitosis) of the cell cycle by specifically binding to tubulin. Therefore, the combined tubulin molecules cannot be polymerized into microtubules. Mitosis is considered to stop in the metaphase, followed by cell death. The example of vinca alkaloids includes but is not limited to vinblastine, vincristine and vinorelbine.

Vinblastine, vinblastine sulfate, is commercially available as an injectable solution as ^VELBAN® . Although it may be indicated as a second-line treatment for various solid tumors, its primary indications are for the treatment of testicular cancer and various lymphomas, including Hodgkin's disease; and lymphocytic and histiocytic lymphomas. Myelosuppression is a dose-limiting side effect of vinblastine.

Vincristine, 22-oxo-vinblastine sulfate, is commercially available as an injection solution as ^ONCOVIN® . Vincristine is indicated for the treatment of acute leukemias and has also found use in treatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas. Alopecia and neurological effects are the most common side effects of vincristine, and myelosuppression and gastrointestinal mucositis effects occur to a lesser extent.

Vinorelbine, 3',4'-didehydro-4'-deoxy-C'-norvincaleukoblastine [R-(R*,R*)-2,3-dihydroxysuccinate (1:2) (salt)], is commercially available as vinorelbine tartrate injection ( ^NAVELBINE® ), a semisynthetic vinca alkaloid. Vinorelbine is indicated for the treatment of various solid tumors, particularly non-small cell lung cancer, advanced breast cancer, and hormone-refractory prostate cancer, as a single agent or in combination with other chemotherapeutic agents such as cisplatin. Myelosuppression is the most common dose-limiting side effect of vinorelbine.

Platinum coordination complexes are non-stage specific anticancer agents that interact with DNA. Platinum complexes enter tumor cells, undergo hydration and form intra- and inter-strand crosslinks with DNA, causing biological effects that are unfavorable to tumors. Examples of platinum coordination complexes include, but are not limited to, oxaliplatin, cisplatin, and carboplatin.

Cisplatin, cis-dichlorodiamine platinum, is commercially available as an injectable solution as ^PLATINOL® . Cisplatin is primarily indicated for the treatment of metastatic testicular and ovarian cancer and advanced bladder cancer.

Carboplatin, diamine[1,1-cyclobutane-dicarboxylate (2-)-O,O']platinum, is commercially available as an injection as ^PARAPLATIN® . Carboplatin is primarily indicated for first-line and second-line treatment of advanced ovarian cancer.

Alkylating agents are non-phasic anticancer specific agents and strong electrophiles. Typically, alkylating agents form covalent bonds with DNA through nucleophilic moieties such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl and imidazole groups of DNA molecules by alkylation. Such alkylation destroys nucleic acid function and causes cell death. Examples of alkylating agents include, but are not limited to, nitrogen mustards such as cyclophosphamide, melphalan and chlorambucil; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; and triazenes such as dacarbazine.

Cyclophosphamide, 2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorinane 2-oxide monohydrate, is commercially available as an injectable solution or tablet as ^CYTOXAN® . Cyclophosphamide is indicated for use as a single agent or in combination with other chemotherapeutic agents in the treatment of malignant lymphomas, multiple myeloma, and leukemias.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injection or tablet as ^ALKERAN® . Melphalan is indicated for the palliative treatment of multiple myeloma and unresectable epithelial ovarian cancer. Myelosuppression is the most common dose-limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]phenylbutyric acid, is commercially available as ^LEUKERAN® tablets. Chlorambucil is indicated for the palliative treatment of chronic lymphatic leukemia and malignant lymphomas such as lymphosarcoma, giant follicular lymphoma, and Hodgkin's disease.

Busulfan, 1,4-butylene glycol dimethanesulfonate, is commercially available as MYLERAN ^{® TABLETS} . Busulfan is indicated for the palliative treatment of chronic myeloid leukemia.

Carmustine, 1,3-bis(2-chloroethyl)-1-nitrosourea, is commercially available as a single vial of lyophilized material as ^BiCNU® . Carmustine is indicated as a single agent or in combination with other agents for the palliative treatment of brain tumors, multiple myeloma, Hodgkin's disease, and non-Hodgkin's lymphoma.

Dacarbazine, 5-(3,3-dimethyl-1-triazene)-imidazole-4-carboxamide, is commercially available as a single vial of the substance as DTIC- ^Dome® . Dacarbazine is indicated for the treatment of metastatic malignant melanoma and for second-line treatment of Hodgkin's disease in combination with other agents.

Antibiotic antineoplastic agents are non-stage specific agents that bind or intercalate into DNA. Typically, such action results in stable DNA complexes or strand breaks that disrupt the normal function of nucleic acids, leading to cell death. Examples of antibiotic antineoplastic agents include, but are not limited to, actinomycins such as dactinomycin, anthracyclines such as daunorubicin and doxorubicin; and bleomycins.

Dactinomycin, also known as actinomycin D, is commercially available in injectable form as ^COSMEGEN® . Dactinomycin is indicated for the treatment of Wilman tumor and rhabdomyosarcoma.

Daunorubicin, (8S-cis)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-naphthacene dione hydrochloride, is commercially available as a liposomal injectable form as ^DAUNOXOME® or as an injectable as ^CERUBIDINE® . Daunorubicin is indicated for inducing remission in the treatment of acute non-lymphocytic leukemia and advanced HIV-associated Kaposi's sarcoma.

Doxorubicin, (8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyso-hexopyranosyl)oxy]-8-glycolyl, 7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-naphthacene dione hydrochloride, is commercially available as an injectable form as ^RUBEX® or ADRIAMYCIN ^RDF® . Doxorubicin is primarily indicated for the treatment of acute lymphoblastic leukemia and acute myeloblastic leukemia, but is also a useful agent in the treatment of some solid tumors and lymphomas.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated from strains of Streptomyces verticillus , is commercially available as ^BLENOXANE® . Bleomycin is indicated as a single agent or in combination with other agents for the palliative treatment of squamous cell carcinoma, lymphoma, and testicular cancer.

Topoisomerase II inhibitors include, but are not limited to, epipodophyllotoxins. Epipodophyllotoxins are stage-specific antitumor agents derived from the mandrake plant. Epipodophyllotoxins typically affect cells in the S and _G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA, causing DNA strand breaks. Strand breaks accumulate, followed by cell death. Examples of epipodophyllotoxins include, but are not limited to, etoposide and teniposide.

Etoposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-ethylidene-β-D-pyranoglucopyranoside], is commercially available as an injectable solution or capsule as ^VePESID® and is commonly known as VP- 16. Etoposide is indicated as a single agent or in combination with other chemotherapeutic agents for the treatment of testicular cancer and non-small cell lung cancer.

Teniposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-thenylidene-β-D-pyranoglucopyranoside], is commercially available as an injection as ^VUMON® and is commonly known as VM-26. Teniposide is indicated as a single agent or in combination with other chemotherapeutic agents for the treatment of acute leukemia in children.

Antimetabolite tumor agents are stage-specific antitumor agents that act on the S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis to limit DNA synthesis. Therefore, the S phase does not proceed, and then the cell dies. Examples of antimetabolite antitumor agents include, but are not limited to, fluorouracil, methotrexate, cytarabine, mercaptopurine, thioguanine, and gemcitabine.

5-fluorouracil, 5-fluoro-2,4-(1H,3H)pyrimidinedione, is commercially available as fluorouracil. Administration of 5-fluorouracil results in inhibition of thymidylate synthesis and is also incorporated into both RNA and DNA. The result is usually cell death. 5-fluorouracil is indicated for the treatment of breast cancer, colon cancer, rectal cancer, gastric cancer, and pancreatic cancer as a single agent or in combination with other chemotherapeutic agents. Other fluoropyrimidine analogs include 5-fluorodeoxyuridine (floxuridine) and 5-fluorodeoxyuridine monophosphate.

Cytarabine, 4-amino-1-β-D-arabinofuranosyl-2(1H)-pyrimidinone, is commercially available as CYTOSAR- ^U® and is commonly referred to as Ara-C. It is believed that cytarabine inhibits DNA chain elongation by terminally incorporating cytarabine into growing DNA chains, while showing cell stage specificity in the S phase. Cytarabine is indicated for the treatment of acute leukemia as a single agent or in combination with other chemotherapeutic agents. Other cytidine analogs include 5-azacytidine and 2',2'-difluorodeoxycytidine (gemcitabine).

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as ^PURINETHOL® . Mercaptopurine inhibits DNA synthesis by a mechanism that has not yet been determined, but shows cell phase specificity in the S phase. Mercaptopurine is indicated for the treatment of acute leukemias as a single agent or in combination with other chemotherapeutic agents. A useful mercaptopurine analog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commercially available as ^TABLOID® . Thioguanine inhibits DNA synthesis by a mechanism that has not yet been determined, but shows cell phase specificity in the S phase. Thioguanine is indicated for the treatment of acute leukemias as a single agent or in combination with other chemotherapeutic agents. Other purine analogs include pentostatin, erythrohydroxynonyladenine (EHNA), fludarabine phosphate, and cladribine.

Gemcitabine, 2'-deoxy-2',2'-difluorocytidine monohydrochloride (β-isomer), is commercially available as ^GEMZAR® . Gemcitabine exhibits cell phase specificity in the S phase by blocking the progression of cells through the G1/S boundary. Gemcitabine is indicated in combination with cisplatin for the treatment of locally advanced non-small cell lung cancer and alone for the treatment of locally advanced pancreatic cancer.

Methotrexate, N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamic acid, is commercially available as methotrexate sodium. Methotrexate exhibits cell stage effects specifically in the S phase by inhibiting DNA synthesis, repair and/or replication by inhibiting dihydrofolate reductase required for the synthesis of purine nucleotides and thymidylate. Methotrexate is indicated as a single agent or in combination with other chemotherapeutic agents for the treatment of choriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, and breast, head, neck, ovarian, and bladder cancers.

Camptothecins, including camptothecin and camptothecin derivatives, are available or under development as inhibitors of topoisomerase I. The cytotoxic activity of camptothecins is believed to be related to their topoisomerase I inhibitory activity. Examples of camptothecins include, but are not limited to, irinotecan, topotecan, and various optical forms of 7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecin described below.

Irinotecan hydrochloride, (4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3',4',6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione hydrochloride, is commercially available as an injection solution ^CAMPTOSAR® . Irinotecan is a derivative of camptothecin that binds to the topoisomerase I-DNA complex along with its active metabolite SN-38. Cytotoxicity is believed to occur as a result of irreparable double-strand breaks caused by the interaction of the topoisomerase I:DNA:irinotecan or SN-38 ternary complex with the replicase. Irinotecan is indicated for the treatment of metastatic cancer of the colon or rectum.

Topotecan hydrochloride, (S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4',6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride, is commercially available as an injection solution ^HYCAMTIN® . Topotecan is a derivative of camptothecin that binds to the topoisomerase I-DNA complex and prevents the relegation of single-strand breaks caused by topoisomerase I in response to torsional tension in the DNA molecule. Topotecan is indicated for second-line treatment of metastatic ovarian cancer and small cell lung cancer.

Hormones and hormone analogs are useful compounds for the treatment of cancer where a relationship exists between the hormone and the growth and/or lack of growth of the cancer. Examples of hormones and hormone analogs that can be used for cancer treatment include, but are not limited to, adrenocortical steroids, such as prednisone and prednisolone, which are useful for treating malignant lymphomas and acute leukemias in children; aminoglutethimide and other aromatase inhibitors, such as anastrozole, letrozole, vorozole, and exemestane, which are useful for treating adrenocortical carcinoma and hormone-dependent breast cancer containing estrogen receptors; progestins, such as megestrol acetate, which are useful for treating hormone-dependent breast cancer and endometrial cancer; estrogens and antiestrogens, such as fulvestrant, flutamide, nilutamide, bicalutamide, cyproterone acetate, and 5α-reductase inhibitors such as finasteride and dutasteride, which are useful for treating prostate cancer and benign prostatic hypertrophy; antiestrogens, such as tamoxifen, toremifene, raloxifene, droloxifene, iodoxyfene, and selective estrogen receptor modulators (SERMS), such as those described in U.S. Pat. No. 5,681,835, 5,877,219 and 6,207,716, which are useful for treating hormone-dependent breast cancer and other susceptible cancers; and gonadotropin-releasing hormone (GnRH) and its analogs, which stimulate the release of luteinizing hormone (LH) and/or follicle-stimulating hormone (FSH), which are useful for treating prostate cancer, for example, LHRH agonists and antagonists such as goserelin acetate and leuprolide.

Signal transduction pathway inhibitors are those inhibitors that block or inhibit the chemical processes that cause changes in cells. As used herein, this change is cell proliferation or differentiation. Signal transduction inhibitors that can be used in the present invention include the following inhibitors: receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domain blockers, serine/threonine kinases, phosphatidylinositol-3 kinases, inositol signal transduction and Ras oncogenes.

Several protein tyrosine kinases catalyze the phosphorylation of specific tyrosyl residues in various proteins involved in regulating cell growth. Such protein tyrosine kinases can be broadly classified as either receptor or non-receptor kinases.

Receptor tyrosine kinases are transmembrane proteins with an extracellular ligand binding domain, a transmembrane domain and a tyrosine kinase domain. Receptor tyrosine kinases are involved in the regulation of cell growth and are commonly referred to as growth factor receptors. Inappropriate or uncontrolled activation of many of these kinases, i.e., abnormal kinase growth factor receptor activity, e.g., by overexpression or mutation, has been shown to lead to uncontrolled cell growth. Therefore, abnormal activity of such kinases has been associated with malignant tissue growth. Therefore, inhibitors of such kinases can provide a method for treating cancer. Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet-derived growth factor receptor (PDGFr), erbB2, erbB4, ret, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor-I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptor, Trk receptor (TrkA, TrkB and TrkC), ephrin (eph) receptor and RET proto-oncogene. Several growth receptor inhibitors are under development and include ligand antagonists, antibodies, tyrosine kinase inhibitors and antisense oligonucleotides. Growth factor receptors and agents that inhibit growth factor receptor function are described, for example, in Kath, John C., Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al., DDT Vol 2, No. 2 February 1997; and Lofts, F.J. et al., "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London.

Tyrosine kinases that are not growth factor receptor kinases are referred to as non-receptor tyrosine kinases. Non-receptor tyrosine kinases that can be used as targets or potential targets of anticancer drugs of the present invention include cSrc, Lck, Fyn, Yes, Jak, cAb1, FAK (focal adhesion kinase), Bruton's tyrosine kinase, and Bcr-Abl. Such non-receptor kinases and agents that inhibit the function of non-receptor tyrosine kinases are described in Sinh, S. and Corey, S.J., (1999) Journal of Hematotherapy and Stem Cell Research 8 (5): 465-80; and Bolen, J.B., Brugge, J.S., (1997) Annual review of Immunology. 15: 371-404.

SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in various enzymes or adaptor proteins, including the PI3-K p85 subunit, Src family kinases, adaptor molecules (Shc, Crk, Nck, Grb2), and Ras-GAP. SH2/SH3 domains as anticancer drug targets are discussed in Smithgall, T.E. (1995), Journal of Pharmacological and Toxicological Methods. 34(3) 125-32.

Serine/threonine kinase inhibitors, including MAP kinase cascade blockers, including blockers of Raf kinase (rafk), mitogen- or extracellular-regulated kinase (MEK) and extracellular-regulated kinase (ERK); and protein kinase C family member blockers, including blockers of PKC (α, β, γ, ε, μ, λ, ι, ζ), IkB kinase family (IKKa, IKKb), PKB family kinases, akt kinase family members and TGFβ receptor kinase. Such serine/threonine kinases and their inhibitors are described in Yamamoto, T., Taya, S., Kaibuchi, K., (1999), Journal of Biochemistry, 126 (5) 799-803; Brodt, P, Samani, A., and Navab, R. (2000), Biochemical Pharmacology, 60. 1101-1107; Massague, J., Weis-Garcia, F. (1996) Cancer Surveys. 27:41-64; Philip, P. A., and Harris, A. L. (1995), Cancer Treatment and Research . 78:3-27, Lackey, K. et al., Bioorganic and Medicinal Chemistry Letters, (10), 2000, 223-226; U.S. Pat. No. 6,268,391; and Martinez-Iacaci, L. et al., Int. J. Cancer (2000), 88(1), 44-52.

Inhibitors of members of the phosphatidylinositol-3 kinase family, including blockers of PI3-kinase, ATM, DNA-PK and Ku, can also be used in the present invention. Such kinases are discussed in Abraham, RT (1996), Current Opinion in Immunology . 8 (3) 412-8; Canman, CE, Lim, DS (1998), Oncogene 17 (25) 3301-3308; Jackson, SP (1997), International Journal of Biochemistry and Cell Biology. 29 (7): 935-8; and Zhong, H. et al., Cancer res, (2000) 60 (6), 1541-1545.

Also useful in the present invention are inositol signaling inhibitors, such as phospholipase C blockers and inositol analogs. Such signaling inhibitors are described in Powis, G., and Kozikowski A., (1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workman and David Kerr, CRC press 1994, London.

Another group of signal transduction pathway inhibitors are inhibitors of Ras oncogenes. Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyltransferase, and CAAX proteases, as well as antisense oligonucleotides, ribozymes, and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild-type mutant ras, thereby acting as antiproliferative agents. Ras oncogene inhibition is discussed in Scharovsky, OG, Rozados, VR, Gervasoni, SI Matar, P. (2000), Journal of Biomedical Science . 7(4) 292-8; Ashby, MN (1998), Current Opinion in Lipidology. 9(2) 99 - 102; and BioChim. Biophys. Acta, (19899) 1423(3): 19-30.

As described above, antibody antagonists of receptor kinase ligand binding can also be used as signal transduction inhibitors. This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases. For example, Imclone C225 EGFR-specific antibody (see Green, MC et al., Monoclonal Antibody Therapy for Solid Tumors, Cancer Treat. Rev., (2000), 26(4), 269-286); ^Herceptin® erbB2 antibody (see Tyrosine Kinase Signalling in Breast cancer:erbB Family Receptor Tyrosine Kinases, Breast cancer Res., 2000, 2(3), 176-183); and 2CB VEGFR2-specific antibody (see Brekken, RA et al., Selective Inhibition of VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumor growth in mice, Cancer Res. (2000) 60, 5117-5124).

Anti-angiogenic therapeutics including non-receptor MEK angiogenesis inhibitors may also be useful. Anti-angiogenic agents such as those that inhibit the action of vascular endothelial growth factor (e.g., the anti-vascular endothelial cell growth factor antibody bevacizumab [Avastin™], and compounds that act through other mechanisms (e.g., linomide, inhibitors of integrin αvβ3 function, endostatin and angiostatin).

Agents for immunotherapy regimens may also be used in combination with compounds of formula (I). Immunotherapeutic methods include, for example, ex vivo and in vivo methods for increasing the immunogenicity of patient tumor cells, such as transfection with cytokines such as interleukin 2, interleukin 4, or granulocyte-macrophage colony stimulating factor, methods for reducing T cell energy, methods using transfected immune cells such as cytokine-transfected dendritic cells, methods using cytokine-transfected tumor cell lines, and methods using anti-idiotypic antibodies.

Therapeutic agents used in pro-apoptotic regimens (eg, bcl-2 antisense oligonucleotides) may also be used in the combinations of the invention.

Cell cycle signal transduction inhibitors inhibit molecules involved in cell cycle control. A family of protein kinases called cyclin-dependent kinases (CDKs) and their interaction with a family of proteins called cyclins control the progression through the eukaryotic cell cycle. Coordinated activation and inactivation of different cyclin/CDK complexes are essential for normal progression through the cell cycle. Several inhibitors of cell cycle signal transduction are under development. For example, examples of cyclin-dependent kinases, including CDK2, CDK4 and CDK6 and their inhibitors are described in, for example, Rosania et al., Exp. Opin. Ther. Patents (2000) 10 (2): 215-230.

In one embodiment, the combination of the present invention comprises a compound of formula (I) or a salt thereof (particularly a pharmaceutically acceptable salt thereof) and at least one anti-tumor agent selected from anti-microtubule agents, platinum coordination complexes, alkylating agents, antibacterial agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine MEK angiogenesis inhibitors, immunotherapeutic agents, pro-apoptotic agents and cell cycle signal transduction inhibitors.

In one embodiment, the combination product of the present invention comprises a compound of formula (I) or a salt thereof (particularly a pharmaceutically acceptable salt thereof) and at least one anti-tumor agent which is an anti-microtubule agent selected from diterpenoids and vinca alkaloids.

In a further embodiment, at least one anti-neoplastic agent is a diterpenoid.In a further embodiment, at least one anti-neoplastic agent is a vinca alkaloid.

In one embodiment, the combination product of the present invention comprises a compound of formula (I) or a salt thereof (particularly a pharmaceutically acceptable salt thereof) and at least one anti-tumor agent which is a platinum coordination complex.

In a further embodiment, at least one antineoplastic agent is paclitaxel, carboplatin or vinorelbine. In a further embodiment, at least one antineoplastic agent is carboplatin. In a further embodiment, at least one antineoplastic agent is vinorelbine. In a further embodiment, at least one antineoplastic agent is paclitaxel. In one embodiment, the combination product of the present invention comprises a compound of formula (I) or a salt, in particular a pharmaceutically acceptable salt thereof, and at least one antineoplastic agent, which is a signal transduction pathway inhibitor.

In a further embodiment, the signal transduction pathway inhibitor is an inhibitor of growth factor receptor kinases VEGFR2, TIE2, PDGFR, BTK, erbB2, EGFr, IGFR-1, TrkA, TrkB, TrkC or c-fms. In a further embodiment, the signal transduction pathway inhibitor is an inhibitor of serine/threonine kinases rafk, akt or PKC-ζ. In a further embodiment, the signal transduction pathway inhibitor is an inhibitor of non-receptor tyrosine kinases selected from kinases of the src family. In a further embodiment, the signal transduction pathway inhibitor is an inhibitor of c-src. In a further embodiment, the signal transduction pathway inhibitor is an inhibitor of Ras oncogenes selected from inhibitors of farnesyl transferase and geranylgeranyl transferase. In a further embodiment, the signal transduction pathway inhibitor is an inhibitor of serine/threonine kinases selected from PI3K.

In a further embodiment, the signal transduction pathway inhibitor is a dual EGFr/erbB2 inhibitor, such as N-{3-chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine.

In one embodiment, the combination product of the present invention comprises a compound of formula (I) or a salt thereof (particularly a pharmaceutically acceptable salt thereof) and at least one anti-tumor agent, wherein the anti-tumor agent is a cell cycle signal transduction inhibitor. In other embodiments, the cell cycle signal transduction inhibitor is an inhibitor of CDK2, CDK4 or CDK6.

Additional examples of other therapeutic agents (eg, anti-neoplastic agents) for combination or co-administration with the compounds of Formula (I) are immunomodulators.

As used herein, "immunomodulator" refers to any substance that affects the immune system, including monoclonal antibodies. Immunomodulators can be used as anti-tumor agents for the treatment of cancer. For example, immunomodulators include, but are not limited to, anti-CTLA-4 antibodies, such as ipilimumab (YERVOY) and anti-PD-1 antibodies (Opdivo/nivolumab and Keytruda/pembrolizumab). Other immunomodulators include, but are not limited to, ICOS antibodies, OX-40 antibodies, PD-L1 antibodies, LAG3 antibodies, TIM-3 antibodies, 41BB antibodies, and GITR antibodies.

Additional examples of other therapeutic agents (anti-tumor agents) for combination or co-administration with the compounds of the present invention are anti-PD-L1 agents. Anti-PD-L1 antibodies and methods for their preparation are known in the art. Such antibodies against PD-L1 may be polyclonal or monoclonal, and/or recombinant and/or humanized. Exemplary PD-L1 antibodies are disclosed in U.S. Patent Nos. 8,217,149, 8,383,796, 8,552,154, 9,212,224, and 8,779,108 and U.S. Patent Application Publication Nos. 20110280877, 2014/0341902, and 20130045201. Additional exemplary antibodies to PD-L1 (also known as CD274 or B7-H1) and methods of use are disclosed in U.S. Patent Nos. 7,943,743, 8,168,179; and 7,595,048, WO2014055897, WO2016007235, and U.S. Patent Application Publication Nos. 20130034559, 20130034559, and 20150274835. PD-L1 antibodies are being developed as immunomodulatory agents or immunomodulators for the treatment of cancer.

In one embodiment, the antibody to PD-L1 is an antibody disclosed in U.S. Patent No. 8,217,149. In another embodiment, the anti-PD-L1 antibody comprises the CDRs of the antibody disclosed in U.S. Patent No. 8,217,149. In another embodiment, the antibody to PD-L1 is an antibody disclosed in U.S. Patent No. 8,779,108. In another embodiment, the anti-PD-L1 antibody comprises the CDRs of the antibody disclosed in U.S. Application No. 8,779,108. In another embodiment, the antibody to PD-L1 is an antibody disclosed in U.S. Patent Application Publication No. 20130045201. In another embodiment, the anti-PD-L1 antibody comprises the CDRs of the antibody disclosed in U.S. Patent Application Publication No. 20130045201. In one embodiment, the anti-PD-L1 antibody is BMS-936559 (MDX-1105) described in WO 2007/005874. In another embodiment, the anti-PD-L1 antibody is MPDL3280A (RG7446). In another embodiment, the anti-PD-L1 antibody is MEDI4736, which is an anti-PD-L1 monoclonal antibody described in WO 2011/066389 and US 2013/034559. In another embodiment, the anti-PD-L1 antibody is TECENTRIQ™ (atezolizumab), which is an anti-PDL1 cancer immunotherapy approved in the United States in May 2016 for a specific type of bladder cancer. In another embodiment, the anti-PD-L1 antibody is YW243.55.S70, which is an anti-PD-L1 described in WO 2010/077634 and U.S. Patent No. 8,217,149. Examples of anti-PD-L1 antibodies useful in the methods of the invention and methods for their preparation are described in PCT patent applications WO 2010/077634, WO 2007/005874, WO 2011/066389, U.S. Pat. No. 8,217,149, and US 2013/034559.

Other examples of mAbs that bind to human PD-L1 and can be used in the methods of treatment, medicines and uses of the present invention are described in WO2013/019906, WO2010/077634A1 and US8383796. Specific anti-human PD-L1 mAbs that can be used as PD-1 antagonists in the methods of treatment, medicines and uses of the present invention include MPDL3280A, BMS-936559, MEDI4736, MSB0010718C.

An additional example of other therapeutic agents (anti-tumor agents) for combination or co-administration with the compounds of the invention is a PD-1 antagonist.

"PD-1 antagonist" means any chemical compound or biological molecule that blocks PD-L1 expressed on cancer cells from binding to PD-1 expressed on immune cells (T cells, B cells or NKT cells) and preferably also blocks PD-L2 expressed on cancer cells from binding to PD-1 expressed by immune cells. Alternative names or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279 and SLEB2 for PD-1; PDCD1L1, PDL1, B7H1, B7-4, CD274 and B7-H for PD-L1; and PDCD1L2, PDL2, B7-DC, Btdc and CD273 for PD-L2. In any embodiment of the aspects or embodiments of the present invention in which a human individual is to be treated, the PD-1 antagonist blocks the binding of human PD-L1 to human PD-1, and preferably blocks the binding of both human PD-L1 and PD-L2 to human PD-1. The human PD-1 amino acid sequence can be found in NCBI Locus No.: NP_005009. The human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively.

PD-1 antagonists that can be used in any aspect of the present invention include monoclonal antibodies (mAbs) or antigen-binding fragments thereof that specifically bind to PD-1 or PD-L1 and preferably specifically bind to human PD-1 or human PD-L1. The mAb can be a human antibody, a humanized antibody, or a chimeric antibody, and can include a human constant region. In some embodiments, the human constant region is selected from IgG1, IgG2, IgG3, and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgG1 or IgG4 constant region. In some embodiments, the antigen-binding fragment is selected from Fab, Fab'-SH, F(ab')2, scFv, and Fv fragments.

Examples of mAbs that bind to human PD-1 and can be used in various aspects and embodiments of the invention are described in US7488802, US7521051, US8008449, US8354509, US8168757, WO2004/004771, WO2004/072286, WO2004/056875 and US2011/0271358.

Specific anti-human PD-1 mAbs that can be used as PD-1 antagonists in any aspects and embodiments of the invention include: MK-3475, a humanized IgG4 mAb having a structure described in WHO Drug Information , Vol. 27, No. 2, pages 161-162 (2013), and which comprises the heavy chain and light chain amino acid sequences shown in Figure 6; nivolumab, a human IgG4 mAb having a structure described in WHO Drug Information , Vol. 27, No. 1, pages 68-69 (2013), and which comprises the heavy chain and light chain amino acid sequences shown in Figure 7; humanized antibodies h409A11, h409A16 and h409A17 (which are described in WO2008/156712) and AMP-514 (which is being developed by Medimmune).

Other PD-1 antagonists that can be used in any aspect and embodiment of the present invention include immunoadhesins that specifically bind to PD-1 and preferably specifically bind to human PD-1, such as fusion proteins containing an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region (such as an Fc region) of an immunoglobulin molecule. Examples of immunoadhesion molecules that specifically bind to PD-1 are described in WO2010/027827 and WO2011/066342. Specific fusion proteins that can be used as PD-1 antagonists in the treatment methods, drugs and uses of the present invention include AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein and binds to human PD-1.

KEYTRUDA/pembrolizumab is an anti-PD-1 antibody marketed by Merck for the treatment of lung cancer. The amino acid sequence and methods of use of pembrolizumab are disclosed in U.S. Patent No. 8,168,757.

Opdivo/Nivolumab is a fully human monoclonal antibody against the negative immunoregulatory human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1/PCD-1) sold by Bristol Myers Squibb, which has immunoenhancing activity. Nivolumab binds to and blocks the activation of PD-1 (Ig superfamily transmembrane protein) through its ligands PD-L1 and PD-L2, leading to the activation of T cells and cell-mediated immune responses against tumor cells or pathogens. Activated PD-1 negatively regulates T cell activation and effector function by inhibiting P13k/Akt pathway activation. Other names for nivolumab include: BMS-936558, MDX-1106 and ONO-4538. The amino acid sequence of nivolumab and methods of use and preparation are disclosed in U.S. Patent No. US 8,008,449.

An additional example of other therapeutic agents (anti-tumor agents) for combination or co-administration with the compounds of formula (I) is an antibody directed against ICOS.

ICOS is a co-stimulatory T cell receptor that has a structural and functional relationship with the CD28/CTLA-4-Ig superfamily (Hutloff, et al., "ICOS is an inducible T-cell co-stimulator structurally and functionally related to CD28", Nature, 397: 263-266 (1999)). ICOS activation occurs through the binding of ICOS-L (B7RP-1/B7-H2). Neither B7-1 nor B7-2 (ligands for CD28 and CTLA4) binds or activates ICOS. However, ICOS-L has been shown to weakly bind both CD28 and CTLA-4 (Yao S et al., "B7-H2 is a costimulatory ligand for CD28 in human", Immunity, 34(5); 729-40 (2011)). The expression of ICOS appears to be limited to T cells. ICOS expression levels vary between different T cell subsets and T cell activation states. ICOS expression has been shown on resting TH17, T follicular helper cells (TFH) and regulatory T (Treg) cells; however, unlike CD28; it is not highly expressed on naive _TH1 and _TH2 effector T cell populations (Paulos CM et al., "The inducible costimulator (ICOS) is critical for the development of human Th17 cells", Sci Transl Med, 2(55); 55ra78 (2010)). After activation by TCR engagement, ICOS expression is highly induced on CD4+ and CD8+ effector T cells (Wakamatsu E, et al., "Convergent and divergent effects of costimulatory molecules in conventional and regulatory CD4+ T cells", Proc Natal Acad Sci USA, 110(3); 1023-8 (2013)).

The CDRs of murine antibodies against human ICOS with agonist activity are shown in PCT/EP2012/055735 (WO2012/131004). Antibodies against ICOS are also disclosed in WO 2008/137915, WO 2010/056804, EP1374902, EP 1374901 and EP 1125585.

Agonist antibodies or ICOS binding proteins to ICOS are disclosed in WO2012/13004, WO 2014/033327, WO2016/120789, US20160215059 and US20160304610. In one embodiment, an agonist antibody to ICOS comprises an ICOS binding protein, or an antigen binding portion thereof, comprising one or more of: a CDRH1 as shown in SEQ ID NO: 1; a CDRH2 as shown in SEQ ID NO: 2; a CDRH3 as shown in SEQ ID NO: 3; a CDRL1 as shown in SEQ ID NO: 4; a CDRL2 as shown in SEQ ID NO: 5 and/or a CDRL3 as shown in SEQ ID NO: 6, or a direct equivalent of each CDR, wherein the direct equivalent has no more than two amino acid substitutions in the CDRs, as disclosed in WO2016/120789 (which is incorporated herein by reference in its entirety). In one embodiment, the ICOS binding protein, or an antigen binding portion thereof, is an agonist antibody to ICOS, comprising a _VH domain (comprising an amino acid sequence having at least 90% identity to the amino acid sequence shown in SEQ ID NO:7) and/or a _VL domain (comprising an amino acid sequence having at least 90% identity to the amino acid sequence shown in SEQ ID NO:8), as described in WO2016/120789, wherein the ICOS binding protein specifically binds to human ICOS. In one embodiment, the ICOS binding protein is an agonist antibody to ICOS, comprising a _VH domain comprising the amino acid sequence shown in SEQ ID NO:7 and a _VL domain comprising the amino acid sequence shown in SEQ ID NO:8, as described in WO2016/120789.

Yervoy (ipilimumab) is a fully human CTLA-4 antibody marketed by Bristol Myers Squibb. The protein structure and methods of use of ipilimumab are described in U.S. Patent Nos. 6,984,720 and 7,605,238.

CD134, also known as OX40, is a member of the TNFR-superfamily of receptors that are not constitutively expressed on resting naive T cells, unlike CD28. OX40 is a secondary co-stimulatory molecule that is expressed 24 to 72 hours after activation; its ligand OX40L is also not expressed on resting antigen-presenting cells, but is expressed after their activation. The expression of OX40 depends on the full activation of T cells; without CD28, the expression of OX40 is delayed and the level is reduced to 1/4. OX-40 antibodies, OX-40 fusion proteins and methods of use thereof are disclosed in U.S. Patent Nos.: US 7,504,101; US 7,758,852; US 7,858,765; US 7,550,140; US 7,960,515; WO2012027328; WO2013028231.

In one embodiment, the OX40 antigen binding protein is one disclosed in WO2012/027328 (PCT/US2011/048752) (International Application Date August 23, 2011). In another embodiment, the antigen binding protein comprises the CDRs of an antibody disclosed in WO2012/027328 (PCT/US2011/048752) (International Application Date August 23, 2011), or CDRs with 90% identity to the disclosed CDR sequences. In a further embodiment, the antigen binding protein comprises VH, VL, or both of an antibody disclosed in WO2012/027328 (PCT/US2011/048752) (International Application Date August 23, 2011), or VH or VL with 90% identity to the disclosed VH or VL sequences.

In another embodiment, the OX40 antigen binding protein is disclosed in WO2013/028231 (PCT/US2012/024570) (International Application Date February 9, 2012, which is incorporated herein by reference in its entirety). In another embodiment, the antigen binding protein comprises the CDRs of the antibodies disclosed in WO2013/028231 (PCT/US2012/024570) (International Application Date February 9, 2012), or CDRs with 90% identity to the disclosed CDR sequences. In a further embodiment, the antigen binding protein comprises VH, VL, or both of the antibodies disclosed in WO2013/028231 (PCT/US2012/024570) (International Application Date February 9, 2012), or VH or VL with 90% identity to the disclosed VH or VL sequences. In one embodiment, the OX40 antigen binding protein is an isolated agonist antibody against OX40 comprising a light chain variable region having a sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 10 as shown in WO2013/028231 and a heavy chain variable region having a sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 4 as shown in WO2013/028231. In one embodiment, the OX40 antigen binding protein is an isolated antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10 as shown in WO2013/028231 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4 as shown in WO2013/028231.

Therefore, in one embodiment, a method of treating a person in need is provided, comprising administering a compound of formula (I) or a salt thereof and at least one immunomodulator. In one embodiment, the immunomodulator is selected from an ICOS agonist antibody, an OX-40 antibody, or a PD-1 antibody. In one embodiment, the person has cancer. Also provided herein is the use of a compound of formula (I) or a salt thereof in combination with at least one immunomodulator for treating a person in need.

An additional example of other therapeutic agents for use or co-administration in combination with the compounds of formula (I) or salts thereof are immunostimulants.

As used herein, "immunostimulator" refers to any agent that can stimulate the immune system. As used herein, immunostimulatory agents include, but are not limited to, vaccine adjuvants such as Toll-like receptor agonists, T-cell checkpoint blockers such as mAbs to PD-1 and CTL4, and T-cell checkpoint agonists such as agonist mAbs to OX-40 and ICOS.

As used herein, the term "Toll-like receptor" (or "TLR") refers to a member or fragment of the Toll-like receptor protein family that senses microbial products and/or initiates an adaptive immune response. In one embodiment, TLR activates dendritic cells (DCs). Toll-like receptors (TLRs) are a family of pattern recognition receptors that were originally identified as sensors of the innate immune system that recognize microbial pathogens. TLRs recognize different structures in microorganisms, often referred to as "PAMPs" (pathogen-associated molecular patterns). Ligands that bind to TLRs trigger a cascade of intracellular signal transduction pathways that induce the production of factors involved in inflammation and immunity. In humans, ten TLRs have been identified. TLRs expressed on the cell surface include TLR-1, -2, -4, -5, and -6, while TLR-3, -7/8, and -9 express ER compartments. Human DC subsets can be identified based on different TLR expression patterns. As an example, the myeloid or "conventional" subpopulation (mDC) of DC expresses TLR 1-8 when stimulated, and produces a cascade of activation markers (e.g., CD80, CD86, class I and class II MHC, CCR7), proinflammatory cytokines and chemokines. The result of this stimulation and the resulting expression is antigen-specific CD4+ and CD8+T cell priming. These DCs acquire the enhanced ability to ingest antigens and present them to T cells in an appropriate form. In contrast, the plasmacytoid subpopulation (pDC) of DC only expresses TLR7 and TLR9 after activation, resulting in activated NK cells and T cells. Because the tumor cells in death may adversely affect DC function, it has been suggested that activating DC with TLR agonists may be beneficial in eliciting anti-tumor immunity in immunotherapy methods for treating cancer. It has also been suggested that the successful treatment of breast cancer using radiation and chemotherapy requires TLR4 activation.

TLR agonists known in the art and useful in the present invention include, but are not limited to, the following: Pam3Cys, a TLR1/2 agonist; CFA, a TLR2 agonist; MALP2, a TLR2 agonist; Pam2Cys, a TLR2 agonist; FSL-I, a TLR-2 agonist; Hib-OMPC, a TLR-2 agonist; polyinosinic:polycytidylic acid (poly I:C), a TLR3 agonist; polyadenosine- Polyuridylic acid (poly-AU), a TLR3 agonist; polyinosinic-polycytidylic acid (HIltonol) stabilized with poly-L-lysine and carboxymethylcellulose, a TLR3 agonist; bacterial flagellin, a TLR5 agonist; imiquimod, a TLR7 agonist; resiquimod, a TLR7/8 agonist; loxoribine, a TLR7/8 agonist; and unmethylated CpG dinucleotide (CpG-ODN), a TLR9 agonist.

Additional TLR agonists known in the art and useful in the present invention also include, but are not limited to, aminoalkyl aminoglucosyl phosphates (AGPs), which bind to the TLR4 receptor and are known to be used as vaccine adjuvants and immunostimulants for stimulating cytokine production in immunized animals, activating macrophages, promoting innate immune responses, and enhancing antibody production. An example of a naturally occurring TLR4 agonist is bacterial lipopolysaccharide. Suitably, the TLR4 agonist is a non-toxic derivative of lipid A. An example of a semi-synthetic non-toxic derivative of a lipid A TLR4 agonist is monophosphoryl lipid A, and particularly 3-de-O-acylated monophosphoryl lipid A (3D-MPL). 3D-MPL is sold by GlaxoSmithKline Biologicals S.A under the name MPL. AGP and its immunomodulatory effect via TLR4 are disclosed in patent publications, such as WO2006/016997, WO 2001/090129 and/or U.S. Patent No. 6,113,918, and have been reported in the literature. Additional AGP derivatives are disclosed in U.S. Patent No. 7,129,219, U.S. Patent No. 6,525,028 and U.S. Patent No. 6,911,434. Some AGPs serve as agonists of TLR4, while others are considered to be TLR4 antagonists.

In one embodiment, the immunostimulant used in combination with the compounds of the invention is a TLR4 agonist. In one embodiment, the TLR4 agonists are referred to as CRX-601 and CRX-527. Their structures are shown below:

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In addition, another preferred embodiment employs the TLR4 agonist CRX 547 having the structure shown.

Still other embodiments include AGPs such as CRX 602 or CRX 526, which provide increased stability for AGPs having shorter secondary acyl or alkyl chains.

Therefore, in one embodiment, a method for treating a person in need is provided, comprising administering a compound of formula (I) or a salt thereof and at least one immunostimulant. In one embodiment, the immunostimulant is a TLR4 agonist. In one embodiment, the immunostimulant is AGP. In yet another embodiment, the TLR4 agonist is selected from a compound of formula CRX-601, CRX-527, CRX-547, CRX-602 or CRX-526. In one embodiment, the person has cancer. Also provided herein is the use of a combination of a compound of formula (I) or a salt thereof and at least one immunostimulant for treating a person in need.

In addition to the immunostimulants described above, the compositions of the present invention may further comprise other therapeutic agents that, due to their adjuvant properties, may act to stimulate the immune system to respond to cancer antigens present on inactivated tumor cells. Such adjuvants include, but are not limited to, lipids, liposomes, inactivated bacteria (e.g., inactivated or attenuated Listeria monocytogenes) that induce innate immunity, compositions that mediate innate immune activation via (NOD)-like receptors (NLDs), retinoic acid-induced gene (RIG)-I-like receptors (RLRs) and/or C-type lectin receptors (CLRs). Examples of PAMPs include lipoproteins, lipopolypeptides, peptidoglycans, yeast polysaccharides, neisseria porins, flagellins, profillin, galactocerimide, muramyl dipeptides. Peptidoglycans, lipoproteins, and lipoteichoic acid are Gram-positive cell wall components. Lipopolysaccharides are expressed by most bacteria, of which MPL is an example. Flagellin refers to the structural component of bacterial flagella secreted by pathogenic and commensal bacteria. rt.-Galactosylceramide (rt.-GalCer) is an activator of natural killer T (NKT) cells. Muramyl dipeptide is a biologically active peptidoglycan motif common to all bacteria.

Due to their adjuvant properties, TLR agonists are preferably used in combination with other vaccines, adjuvants and/or immunomodulators, and can be combined in various combination products. Therefore, in some embodiments, the compounds of formula (I) described herein that bind to STING and induce STING-dependent TBKI activation and the inactivated tumor cells described herein that express and secrete one or more cytokines that stimulate DC induction, recruitment and/or maturation (as described herein) can be administered together with one or more TLR agonists for therapeutic purposes.

Indoleamine 2,3-dioxygenase 1 (IDO1) is a key immunosuppressive enzyme that regulates anti-tumor immune responses by promoting the generation of regulatory T cells and blocking effector T cell activation, thereby promoting tumor growth by allowing cancer cells to avoid immune surveillance. (Lemos H et al., Cancer Res. 2016 Apr 15; 76(8): 2076-81), (Munn DH et al., Trends Immunol. 2016 Mar; 37(3): 193-207). The other active ingredient (antitumor agent) used in combination or co-administered with the compound of formula (I) of the present invention is an IDO inhibitor. Epacadostat, ((Z)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-4-[2-(sulfamoylamino)ethylamino]-1,2,5-oxadiazole-3-carboxamidine) is a highly potent and selective oral inhibitor of the IDO1 enzyme that reverses tumor-associated immunosuppression and restores effective anti-tumor immune responses. Epacadostat is disclosed in U.S. Pat. No. 8,034,953.

Additional examples of other therapeutic agents (anti-tumor agents) for combination or co-administration with the compounds of formula (I) are CD73 inhibitors and A2a and A2b adenosine antagonists.

In one embodiment, the compounds of the invention may be used in conjunction with other therapeutic approaches for treating infectious diseases. In particular, antiviral agents and antibacterial agents are contemplated.

The compounds of formula (I) and their pharmaceutically acceptable salts may be used in combination with at least one other therapeutic agent useful for preventing or treating bacterial and viral infections. Examples of such agents include, but are not limited to, polymerase inhibitors such as those disclosed in WO 2004/037818-A1, and those disclosed in WO 2004/037818 and WO 2006/045613; JTK-003, JTK-019, NM-283, HCV-796, R-803, R1728, R1626, and WO 2006/018725, WO 2004/074270, WO 2003/095441, US 2005/0176701, WO 2006/020082, WO 2005/080388, WO 2004/064925, WO 2004/065367, WO 2003/007945, WO 02/04425、WO 2005/014543、WO 2003/000254、EP 1065213、WO 01/47883、WO 2002/057287、WO Those disclosed in 2002/057245 and similar agents; replication inhibitors such as acyclovir, famciclovir, ganciclovir, cidofovir, lamivudine and similar agents; protease inhibitors such as the HIV protease inhibitors saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, brenavir, atazanavir, tipranavir, palinavir, lasinavir and HCV protease inhibitors BILN2061, VX-950, SCH503034; and similar agents; nucleoside and nucleotide reverse transcriptase inhibitors such as zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxil, fozivudine, todoxil, emtricitabine, alovudine, aminodosulf , eficitabine, tenofovir disoproxil fumarate, tenofovir alafenamide fumarate/hemifumarate, and similar agents; non-nucleoside reverse transcriptase inhibitors (including agents with antioxidant activity, such as immunocal, oltipraz, etc.), such as nevirapine, delavirdine, efavirenz, loviramide, immunocal, oltipraz, capravirine, TMC-278, TMC-125, etravirine, rilpivirine, and similar agents; entry inhibitors, such as enfuvirtide (T-20), T-1249, PRO-542, PRO-140, TNX-355, BMS-806, 5-Helix, and similar agents; integrase inhibitors, such as dolutegravir, elvitegravir, raltegravir L-870, 180 and similar agents; budding inhibitors such as PA-344 and PA-457 and similar agents; chemokine receptor inhibitors such as viribviroc (Sch-C), Sch-D, TAK779, maraviroc (UK-427,857), TAK449, and WO 02/74769, WO 2004/054974, WO 2004/055012, WO 2004/055010, WO 2004/055016, WO 2004/055011 and WO 2004/054581 and similar agents; pharmacokinetic enhancers such as cobicistat; neuraminidase inhibitors such as CS-8958, zanamivir, oseltamivir, peramivir and similar agents; ion channel blockers such as amantadine or rimantadine and similar agents; and interfering RNA and antisense oligonucleotides and such as ISIS-14803 and similar agents; antiviral agents of undetermined mechanism of action, such as those disclosed in WO 2005/105761, WO 2003/085375, WO 2006/122011, ribavirin and similar agents.

The compounds of formula (I) and their pharmaceutically acceptable salts may also be used in combination with other therapeutic agents useful for treating Kaposi's sarcoma-associated herpesvirus infection (KSHV and KSHV-associated), including but not limited to chemotherapeutic agents such as bleomycin, vinblastine, vincristine, cyclophosphamide, prednisone, alitretinoin and liposomal anthracyclines such as doxorubicin, daunorubicin, immunotherapeutic agents such as rituximab, tocilizumab, siltuximab and others such as paclitaxel and rapamycin.

In one embodiment of the invention, at least one other therapeutic agent is an antimycobacterial agent or a bactericidal antibiotic. The compounds of formula (I) and their pharmaceutically acceptable salts may also be used in combination with at least one other therapeutic agent that can be used to treat TB infection (Mycobacterium tuberculosis) and tularemia (Francisco tularensis), including but not limited to first-line oral agents isoniazid, rifampicin, pyrazinamide, ethambutol, streptomycin, rifabutin; injectable agents, including kanamycin, amikacin, capreomycin, streptomycin; fluoroquinolones, including levofloxacin, moxifloxacin, ofloxacin; oral antibacterial agents para-aminosalicylic acid, cycloserine, triazone, thioamide, prothionamide; SQ-109, PNU-100480, rifapentine, linezolid, PA-824 AZD5847, gatifloxacin-moxifloxacin, Sirturo (bedaquiline) delamanid (OPC-67683), and drugs with uncertain mechanisms of action in the treatment of drug-resistant TB, including clofazimine, linezolid, amoxicillin/clavulanate thiosemicarbazide imipenem/cilastatin high-dose isoniazid clarithromycin, and ciprofloxacin. The compounds of formula (I) and their pharmaceutically acceptable salts may also be combined with antimycobacterial agents (such as isoniazid (INH), ethambutol ( ^Myambutol® ), rifampicin ( ^Rifadin® ) and pyrazinamide (PZA)); bactericidal antibiotics (such as rifabutin ( ^Mycobutin® ) or rifapentine ( ^Priftin® )), aminoglycosides ( ^{Capreomycin®} ), fluoroquinolones (levofloxacin, moxifloxacin, ofloxacin), thioamides (ethionamide), cyclosporine ( ^Sandimmune® ), para-aminosalicylic acid ( ^Paser® ), cycloserine ( ^Seromycin® ), kanamycin ( ^Kantrex® ), streptomycin, puromycin, capreomycin ( ^Capastat® ), bedaquiline fumarate ( ^Sirturo® ), oxazolidinone (Sutezolid ^... ), PNU-100480 or delamanid (OPC-67683).

The compound of formula (I) and its pharmaceutically acceptable salts can also be used in combination with at least one other therapeutic agent that can be used to treat Chlamydia, including but not limited to azithromycin, doxycycline, erythromycin, levofloxacin, ofloxacin.

The compounds of the invention may also be used in combination with at least one other therapeutic agent useful for treating Plasmodium infection, including but not limited to chloroquine, atovaquone-proguanil, artemether-lumefantrine, mefloquine, quinine, quinidine, doxycycline, clindamycin, artesunate, primaquine.

In the treatment of amyotrophic lateral sclerosis (ALS), a compound of formula (I) or a pharmaceutically acceptable salt thereof may be used in combination with: a glutamate blocker (riluzole (Rilutek®)), quinidine (Nuedexta®), an anticholinergic (amitriptyline®, Artane®, scopolamine patch (Transderm Scop®)), a sympathomimetic (pseudoephedrine), a mucolytic (guaifenesin), or an analgesic (tramadol (Ultram®)); ketorolac (Toradol®); morphine; fentanyl patch (Duragesic®)).

In the treatment of multiple sclerosis, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be used in combination with: corticosteroids (prednisone, methylprednisolone), interferon beta 1-A (Avonex®, Extavia®, Rebif®, Betaseron®), peginterferon beta-1A (Plegridy®), glatiramer acetate (Copaxone®); glatiramer acetate (Glatopa®-generic equivalent of Copaxone); dimethyl fumarate (Tecfidera®); fingolimod (Gilenya®); teriflunomide (Aubagio®); dalfampridine (Ampyra®); daclizumab (Zinbryta); alemtuzumab (Lemtrada®); natalizumab (Tysabri®); or mitoxantrone hydrochloride (Novantrone®).

The compounds of the invention can also be used as adjuvants to improve the immune response and/or reduce the reactogenicity/toxicity to any given antigen in patients, particularly humans, in need thereof. Thus, the compounds of the invention can be used in combination with vaccine compositions to modify, in particular enhance, the immune response, for example by increasing the level or duration of protection and/or allowing a reduction in the antigen dose.

The compound of formula (I) and its pharmaceutically acceptable salt can be used in combination with one or more vaccines or immunogenic antigens for preventing or treating viral infection. Such vaccines or immunogenic antigens include, but are not limited to pathogen-derived proteins or particles such as attenuated viruses, viral particles and viral proteins commonly used as immunogenic substances. Examples of viruses and sources of viral antigens include, but are not limited to, poliovirus, Coronaviridae and Coronavirus, Rhinovirus (all subtypes), Adenovirus (all subtypes), Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis D, Human Papillomavirus (including all subtypes), Rabies virus, Human T-lymphotropic virus (all subtypes), Rubella virus, Mumps virus, Coxsackievirus A (all subtypes), Coxsackievirus B (all subtypes), Human enterovirus, Herpesvirus (including cytomegalovirus), Epstein-Barr virus, Human herpesvirus (all subtypes), Herpes simplex virus, Varicella-zoster virus, Human immunodeficiency virus (HIV) (all subtypes), Epstein-Barr virus, Reovirus (all subtypes), Filovirus (Filovivirus), ruses) (including Marburg virus and Ebola virus (all strains)), Arenaviruses (including Lymphocytic Choriomeningitis virus, Lassa virus, Junin virus and Machupo virus), Arboviruses (including West Nile virus), Dengue virus (all serotypes), Zika virus, Colorado tick fever virus, Sindbis virus, Togaviridae, Flaviviridae, Bunyaviridae, Reoviridae, Rhabdoviridae, Orthomyxoviridae, Poxviruses (including Orthopoxviruses (Variola virus, Monkeypox virus, Vaccinia virus, Vaccinia virus), Yatapoxviruses (Tenapox virus, Yaba monkey tumor virus), Parapoxviruses, Molluscumpox virus), Yellow fever, Hantaviruses (including Hantaan, Seoul, Dobrava, Sin Nombre, Puumala and Dobrava-like Saaremaa), human parainfluenza and influenza viruses (all types), H1N1 influenza and swine influenza viruses, respiratory syncytial virus (all subgroups), rotaviruses (including human rotavirus A-E, bovine rotavirus, macaque rotavirus), polyomaviruses (including simian virus 40, JC virus, BK virus, Colorado tick fever virus (Coltiviruses), Eichhornia virus, Calicivirus), and Parvoviridae (including Dependovirus, Parvovirus and Erythrovirus).

On the other hand, the present invention provides a method for curing HIV, comprising administering the compounds of the present invention to a subject. Patient "cure" ("cure" or "curing") is used to indicate the elimination, cessation, stop or termination of human immunodeficiency virus or symptoms, or the elimination, cessation, stop or termination of the progression of the symptoms or virus for a certain period of time. As an example, in one embodiment, "cure" refers to therapeutic administration or combined administration, which alone or in combination with one or more agents to induce and maintain sustained viral control of human immunodeficiency virus (undetectable levels of plasma viremia obtained by, for example, polymerase chain reaction (PCR) test, bDNA (branched DNA) test or NASBA (nucleic acid sequence-based amplification) test) without any other therapeutic intervention after a minimum of, for example, one or two years. The above-mentioned PCR, bDNA and NASBA tests are performed using techniques known and familiar to those skilled in the art. As an example, the elimination, cessation, stop or termination of human immunodeficiency virus or symptoms, or the elimination, cessation, stop or termination of the progression of the symptoms or virus may last for at least two years.

In another embodiment of the present invention, there is provided a compound of the present invention for use in curing HIV infection.

In another embodiment of the present invention, there is provided the use of a compound of the present invention in the preparation of a medicament for curing HIV infection.

In another aspect, a combination product is provided, which comprises a compound of the invention and one or more additional pharmaceutically active agents for HIV. Such compounds and agents may be present in a pharmaceutical preparation or composition. Therefore, the present invention also includes a method for treating, curing and/or preventing HIV infection in a subject, i.e., administering to the subject a combination product (or a pharmaceutical preparation or composition thereof) comprising a compound of the invention and one or more additional pharmaceutically active agents for HIV.

In such an embodiment, the one or more additional active agents against HIV are selected from zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxil, fozivudine, todoxil), emtricitabine, alovudine, amdosovir, efcitabine, nevirapine, delavirdine, efavirenz, loviride, immunocal, oltipraz, caprovirine, lersivirine, GSK2248761, TMC-278, TMC-125, etravirine, saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, becanavir, darunavir, atazanavir, tipranavir, palinavir, lasinavir, enfuvirtide, T-20, T-1249, PRO-542, PRO-140, TNX-355, BMS-806, BMS-663068 and BMS-626529, 5-Helix, raltegravir, elvitegravir , dolutegravir, cabotegrair, vicriviroc (Sch-C), Sch-D, TAK779, maraviroc, TAK449, didanosine, tenofovir, lopinavir, and darunavir.

Therefore, the compounds of the present invention and one or more other pharmaceutically active agents can be administered together or separately, and when administered separately, they can be administered simultaneously or sequentially in any order. The amount of the compounds of the present invention and the one or more other pharmaceutically active agents and the relative time of administration can be selected to achieve the desired combined therapeutic effect. The combined administration of the compounds of the present invention and other therapeutic agents can be combined by concomitant administration in the following manner: (1) a single pharmaceutical composition comprising two compounds; or (2) a separate pharmaceutical composition each comprising one of the compounds. Alternatively, the combination product can be administered separately in a sequential manner, wherein one therapeutic agent is administered first, then the other second, and vice versa. Such sequential administration can be close in time or distant in time. The amount of the compounds of the present invention and one or more other pharmaceutically active agents for HIV and the relative time of administration are selected to achieve the desired combined therapeutic effect.

In addition, the compounds of the present invention can be used in combination with one or more other agents that can be used to prevent, treat or cure HIV. Examples of such agents include: nucleotide reverse transcriptase inhibitors such as zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxil, fozivudine, todoxil, emtricitabine, alovudine, amide, efavirenz, TDF, TAF and similar agents; non-nucleotide reverse transcriptase inhibitors (including agents with antioxidant activity, such as imipenem, oltipraz, etc.) such as nevirapine, delavirdine, efavirenz, loviride, imipenem, oltipraz, caprovirine, lersivirine, GSK2248761, TMC-278, TMC-125, etravirine, and similar agents; protease inhibitors such as saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, becanavir, darunavir, atazanavir, tipranavir, palinavir, lasinavir, and similar agents; integrase inhibitors such as raltegravir, elvitegravir, bitravir, dolutegravir, cabotegravir and similar agents; maturation inhibitors such as PA-344 and PA-457 and similar agents; and GSK2838232. CXCR4 and/or CCR5 inhibitors such as vicriviroc (Sch-C), Sch-D, TAK779, maraviroc (UK 427,857), TAK449, and those disclosed in WO02/74769, PCT/US03/39644, PCT/US03/39975, PCT/US03/39619, PCT/US03/39618, PCT/US03/39740, and PCT/US03/39732, and similar agents. Other examples of the compounds of the present invention that can be used in combination with one or more agents useful for preventing or treating HIV are listed in Table A.

Table A

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The scope of the combination products of the compounds of the present invention and HIV agents is not limited to those described above, and in principle also includes any combination products with any pharmaceutical composition that can be used to cure, treat and/or prevent HIV. As described, in such a combination product, the compounds of the present invention and other HIV agents can be administered individually or in combination. In addition, one agent can be administered before, simultaneously with or after the administration of one or more other agents.

The present invention can be used in combination with one or more agents that can be used as pharmacological enhancers, in the presence or absence of additional compounds for preventing or treating HIV. Examples of such pharmacological enhancers (or pharmacokinetic enhancers) include, but are not limited to, ritonavir, GS-9350, and SPI-452.

Ritonavir is 10-hydroxy-2-methyl-5-(1-methylethyl)-1-1[2-(1-methylethyl)-4-thiazolyl]-3,6-dioxo-8,11-bis(phenylmethyl)-2,4,7,12-tetraazatridecane-13-oic acid, 5-thiazolylmethyl ester [5S-(5S*,8R*,10R*,11R*)] and is available from Abbott Laboratories of Abbott Park, Illinois as Norvir. Ritonavir is an HIV protease inhibitor indicated for use with other antiretroviral agents in the treatment of HIV infection. Ritonavir also inhibits P450-mediated drug metabolism and the P-glycoprotein (Pgp) cellular transport system, resulting in increased concentrations of the active compound in the body.

GS-9350 is a compound being developed by Gilead Sciences of Foster City California as a drug enhancer.

SPI-452 is a compound being developed by Sequoia Pharmaceuticals of Gaithersburg, Maryland as a drug enhancer.

The aforementioned additional therapeutic agents, when used in combination with the compounds of the invention, may be used, for example, in those amounts specified in the Physicians' Desk Reference (PDR) or in other amounts as determined by one of ordinary skill in the art.

In another embodiment of the invention, there is provided a method of treating HIV infection in a subject comprising administering to the subject a combination product as described herein.

In another embodiment of the invention, there is provided a method of curing an HIV infection in a subject comprising administering to the subject a combination product as described herein.

In another embodiment of the invention, there is provided a method of preventing HIV infection in a subject comprising administering to the subject a combination product as described herein.

In another embodiment of the invention there is provided a combination product as described herein for use as a medicament for the treatment of HIV.

In another embodiment of the present invention there is provided a combination product as described herein for use as a medicament for the prevention of HIV.

In another embodiment of the present invention there is provided a combination product as described herein for use as a medicament for curing HIV.

In another embodiment of the present invention there is provided a combination product as described herein for use in the treatment of HIV infection.

In another embodiment of the present invention there is provided a combination product as described herein for use in the prevention of HIV infection.

In another embodiment of the present invention there is provided a combination product as described herein for use in curing HIV infection.

In another embodiment of the present invention there is provided the use of a combination product as described herein in the preparation of a medicament for the treatment of HIV infection.

In another embodiment of the present invention there is provided the use of a combination product as described herein in the preparation of a medicament for the prevention of HIV infection.

In another embodiment of the present invention there is provided the use of a combination product as described herein in the preparation of a medicament for curing HIV infection.

Therefore, the present invention provides an immunogenic composition comprising an antigen or antigen composition and a compound of formula (I) or a pharmaceutically acceptable salt thereof. Further provided is a vaccine composition comprising an antigen or antigen composition and a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The compounds of formula (I) and their pharmaceutically acceptable salts may also be used in combination with at least one other therapeutic agent useful for preventing or treating viral infections, such as immunotherapy (e.g., interferons or other cytokines/chemokines, cytokine/chemokine receptor modulators, cytokine agonists or antagonists, and similar agents); and therapeutic vaccines, anti-fibrotic agents, anti-inflammatory agents such as corticosteroids or NSAIDs (non-steroidal anti-inflammatory agents), and similar agents.

Compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with other anti-inflammatory agents, including oral or topical corticosteroids, anti-TNF agents, 5-aminosalicylic acid and mesalamine preparations, hydroxychloroquine, thiopurines, methotrexate, cyclophosphamide, cyclosporin, calcineurin inhibitors, mycophenolic acid, mTOR inhibitors, JAK inhibitors, Syk inhibitors, RIPK1 and RIPK2 inhibitors, anti-inflammatory biologics, including anti-IL6 biologics, anti-IL1 agents, anti-IL17 biologics, anti-CD22, anti-integrin agents, anti-IFNa, anti-CD20 or CD4 biologics, and other cytokine inhibitors or biologics directed against T cell or B cell receptors or interleukins.

For example, in the treatment of systemic lupus erythematosus and related lupus disorders, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with at least one other therapeutic agent, including corticosteroids (such as prednisolone ( ^Delatsone® , Orapred, Millipred, Omnipred, Econopred, Flo-Pred), immunosuppressants (such as methotrexate ( ^Rhuematrex® , ^Trexall® )), dexamethasone ( ^Decadron® , ^Solurex® ), mycophenolate mofetil ( ^Cellcept® ), ^tacrolimus® , ^Sirolimus® ), B cell therapy (belimumab (Benlysta® ⁾ , B cell inhibitors ( ^Atacicept® , Apratuzumab® ^), (anti-CD22), SBI-087 (anti-CD20), anti-BAFF antibodies (LY2127399, A623), Velcade ^® ), azathioprine (Azasan ^® , Imuran ^® ), triamcinolone (Clinacort ^® , Kenalog-10 ^® ), hydroxychloroquine (Plaquenil ^® ), thalidomide (Immunoprin ^® , Contergan ^® ), immunoglobulin therapy (HyQiva ^® , Flebogamma ^® , Gamunex ^® , Privigen ^® , Gammagard ^® ), anti-interferon-α therapy (Rontalizumab ^® , sifalimumab ^® , AGS-009 ^® , IFN Kinoid), anti-interferon receptor (IFNR) (Anifrolumab®), TLR7 and TLR9 blockers (IMO-3100), anti-cytokine therapies (anti-IL6 (CNTO-136), anti-interferon-γ (AMG811), immunomodulatory therapies (Lupuzor™, abatacept, Orencia ^® , AMG557, laquinimod, paquinimod, leflunomide, anti-ICOS (Medi-570), anti-CD40 ligand antibody (CDP7657)) and/or platelet aggregation inhibitors (aspirin).

In the treatment of Sjögren's syndrome, compounds that modulate STING, in particular compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with anti-rheumatic agents (hydroxychloroquine and ^Plaquenil® , ^Ridaura® , ^Kineret® ), cholinergic agonists ( ^Salagen® , ^Evoxac® ), JAK inhibitors ( ^Xelijanz® and anti-TNFα therapeutics (Remicade® ^, Humira® ^{, Enbrel®} , ^Cimzia® , ^Simponi® ).

In the treatment of vasculitis and diseases with inflammation of small or medium-sized vessels, compounds that modulate STING, in particular compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with alkylating agents (cyclophosphamide, ^Cytoxan® ), anti-rheumatic anti-CD20 antibodies ( ^Rituxan® , ^Rituximab® ) and anti-TNFα inhibitors (Etanrcept® ⁾ .

In the treatment of psoriasis, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with isibetamab, tildrakizumab, secukinumab, alefacept, calcipotriol and betamethasone dipropionate, prednisone, tazarotene topical gel, methotrexate, cyclosporine, fumaric acid, acitretin, phototherapy (UVA, UVB), psoralen, coal tar, TNF inhibitors (etanercept, infliximab, adalimumab, certolizumab pegol), PDE-4 inhibitors (apremilast), JAK inhibitors (tofacitinib), IL 12/23 (ustekinumab), IL17 (secukinumab, isibetamab, brodalumab (AMG-827)), IL23 (tildrakizumab (MK-3222), guselkumab, itrazumab, biosimilar infliximab (Remsima (Inflectra®), Sandoz GP 11111), biosimilar rituximab (CT-P10 (Mabthera®), PF-05280586 (MabThera®)), biosimilar etanercept (CHS-2014), biosimilar adalimumab (GP-2017), M-518101 topical vitamin D; Maruho GK-664 or CT-327 (topical tropomyosin receptor kinase A), CF-101, secukinumab (AIN457), or dimethyl fumarate LAS-41008.

In the treatment of rheumatoid arthritis, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with: tocilizumab, DMARDs (methotrexate, hydroxychloroquine, sulfasalazine, leflunomide), sulfasalazine delayed release, certolizumab pegol, ibuprofen, naproxen sodium, adalimumab, anakinra (Kineret); etodolac, naproxen sodium, abatacept, prednisone, infliximab, golimumab, rofecoxib, tofacitinib, methotrexate, selective JAK1 & JAK2 inhibitors (baracitinib), antisense oligonucleotides (alicafosen), biosimilars of infliximab (Remsima (Inflectra®)), GS-071 infliximab (Aprogen), SB2 infliximab, PF-06438179 infliximab, GP11111, Biosimilars of rituximab (CT-P10 Rituximab Celltrion), BI-695500, GP-2013, PF-05280586, biosimilars of etanercept (etanercept SB4 (Brenzys™), Benepali®; CHS-0214 etanercept, GP-2015, biosimilars of adalimumab (ABP-501 adalimumab, BI-695501, Samsung SB5, GP-2017, PF-06410293, Momenta M923), or biosimilars of abatacept (M834).

In another embodiment, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered to a patient in need thereof in combination with at least one other therapy and/or at least one other active therapeutic agent that is considered standard of care (U.S. Department of Health and Human Services, Agency for Healthcare Research and Quality, National Guideline Clearinghouse, https://www.guideline.gov/ and World Health Organization, http://www.who.int/management/quality/standards/en/) for any of the diseases and/or conditions described herein.

In another embodiment, a compound that modulates STING, particularly a compound of formula (I) or a pharmaceutically acceptable salt thereof, can be administered to a patient in need thereof in combination with at least one other therapy, for example, in combination with UVA and/or UVB phototherapy indicated for the treatment of psoriasis.

In another embodiment, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered to a patient in need thereof in combination with at least one other active therapeutic agent for an indication described herein, wherein the at least one other active therapeutic agent is: corticosteroids [orally, topically, by injection or as a suppository; prednisone, methylprednisolone, prednisolone, budesonide, betamethasone, dexamethasone, hydrocortisone, triamcinolone acetonide, fluticasone (fluticasone furoate, fluticasone propionate), flurandrenolide (flurane acetonide, flurandrenolide), fluocinolone acetonide, clobetasol (clobetasol propionate)], anti-TNF biologics (etanercept, adalimumab, infliximab, certolizumab or golimumab), PDE-4 inhibitors (apremilast), 5-aminosalicylic acids (mesalamine/mesalamine; sulfasalazine, balsalazide), DMARDs (Disease-modifying antirheumatic drugs: methotrexate, hydroxychloroquine, sulfasalazine, leflunomide), thiopurines (azathioprine, mercaptopurine), JAK inhibitors (tofacitinib), NSAIDs (aspirin, acetaminophen, ibuprofen, naproxen (naproxen sodium), etodolac, celecoxib, diclofenac, meloxicam), anti-IL6 biologics (tocilizumab), anti-IL1 biologics (anakinra, canakinumab, rilonacept), anti-IL12 or IL23 biologics (ustekinumab, risankizumab, guselkumab, tildrakizumab), anti-CD6 biologics (itrazumab), anti-integrin agents (natalizumab (Tysabri®), etrolizumab), anti-IL17 biologics (secukinumab, isibetamab, brodalumab), anti-CD22 biologics (epazumab), anti-CD20 biologics (rituximab, ofatumumab), anti-CD20 or CD4 biologics and other cytokine inhibitors or biologics T-cell or B-cell receptors or interleukins, calcineurin inhibitors (cyclosporine, pimecrolimus, tacrolimus), acitretin, fumaric acid, dimethyl fumarate, cyclophosphamide, cyclosporine (or cyclosporine), methotrexate, mycophenolic acid (or mycophenolate mofetil), topical vitamin D (calcipotriol or calcipotriol), mTOR inhibitors (temsirolimus, everolimus), Syk inhibitors (fotantinib), anti-IFNa biologics (sifalimumab) or retinoids (tazarotene). Other examples of suitable biologics include abatacept, belimumab and alicafosen.

In one embodiment of the invention, the at least one additional therapeutic agent is selected from an inhaled corticosteroid, a long-acting beta agonist, a combination of an inhaled corticosteroid and a long-acting beta agonist, a short-acting beta agonist, a leukotriene modifier, an anti-IgE, a methylxanthine bronchodilator, a mast cell inhibitor, and a long-acting muscarinic antagonist. For example, in the treatment of asthma, compounds that inhibit STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, may be administered in combination with an inhaled corticosteroid (ICS) such as fluticasone propionate ( ^Flovent® ), beclomethasone propionate ( ^QVAR® ), budesonide ( ^Pulmicort ), trimcinolone acetonide (Azmacort®), flunisolide ( ^Aerobid® ), mometasone furoate ( ^{Asmanex® Twisthaler®} ) or ciclesonide ( ^Alvesco® ), a long-acting beta agonist (LABA) such as formoterol fumarate ( ^Foradil® ), salmeterol xinafoate ( ^Serevent® ), a combination of ICS and a LABA such as fluticasone furoate and vilanterol (Breo ^Ellipta® ), a formoterol/budesonide inhaler (Symbicort®), or a combination of an inhaled corticosteroid ( ^ICS) . ), beclomethasone propionate/formoterol (Inuvair ^® ) and fluticasone propionate/salmeterol (Advair ^® ), short-acting beta agonists (SABAs) such as albuterol sulfate (ProAir ^® , Proventil HFA ^® , Ventolin HFA ^® , AccuNeb ^® inhalation solution), levosalbutamol tartrate (Xopenex ^® HFA), ipratropium bromide/albuterol (Combivent ^® Respimat ^® ), ipratropium bromide (Atrovent ^® HFA), leukotriene modifiers such as montelukast sodium (Singulair ^® ), zafirlukast (Accolate ^® ), or zileuton (Zyflo ^® ) and anti-IgE such as omalizumab (Xolair ^® )), methylxanthine bronchodilators such as theophylline (Accurbron ^® , Aerolate ^® , Aquaphyllin ^® , Asbron ^® , Bronkodyl ^® , Duraphyl ^® , Elixicon ^® , Elixomin ^® , Elixophyllin ^® , Labid ^® , Lanophyllin ^® , Quibron-T ^® , Slo-Bid ^® , Slo-Phyllin ^® , Somophyllin ^® , Sustaire ^® , Synophyllin late ^® , T-Phyll ^® , Theo-24 ^® , Theo-Dur ^® , Theobid ^® , Theochron ^® , Theoclear ^® , Theolair ^® , Theolixir ^® , Theophyl ^® , Theovent ^® , Uni-dur ^® , Uniphyl ^® ), mast cell inhibitors such as cromulyn sodium (Nasalcrom ^® ) and nedocromil sodium (Ti lade ^® )), long-acting muscarinic antagonists (LAMA) such as mometasone furoate/formoterol fumarate dihydrate (Dulera ^® )).

Other agents that may be useful in combination therapies for the treatment of asthma include protein tyrosine kinase inhibitors (masitinib), CRTH2/D-prostanoid receptor antagonists (AMG 853), indacaterol (Arcapta ^® Neohaler ^® ), epinephrine inhalation aerosol (E004), fluticasone furoate/fluticasone propionate, vinanterol inhalation/fluticasone furoate powder (Relovair™), fluticasone propionate/formoterol fumarate dehydrate (Flutiform ^® ), reslizumab, albuterol dry powder inhaler, tiotropium (Spiriva ^® HandiHaler ^® ), formoterol/budesonide (Symbicort ^® SMART ^® ), fluticasone furoate (Veramyst ^® ), Vectura's VR506, levitra (RG3637), and a combination of phosphodiesterase (PDE)-3 and (PDE)-4 inhibitors (RPL554).

In one embodiment of the invention, the at least one additional therapeutic agent is selected from a long-acting beta agonist, a long-acting inhaled anticholinergic or muscarinic antagonist, a phosphodiesterase inhibitor, an inhaled corticosteroid, a combination of a long-acting beta agonist, a short-acting beta agonist, and an inhaled corticosteroid. For example, in the treatment of COPD, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with a LABA (such as salmeterol xinafoate (Serevent), umeclidinium/vilanterol (Anoro ^Ellipta® ), umeclidinium (Incruse ^Ellipta® ), formoterol tartrate (Brovana®), formoterol fumarate inhalation powder ( ^Foradil® ), indacaterol maleate ( ^{Arcapta® Neohaler®} ), or fluticasone propionate/formoterol fumarate dehydrate (Flutiform®)), a long-acting ^inhaled anticholinergic (or muscarinic antagonist, such as tiotropium ( ^{Spiriva® )} and aclidinium ( ^{Tudorza® Pressair®} ), a phosphodiesterase (PDE-r) inhibitor (such as roflumilast, Daliresp® ⁾ , or a combination thereof. ), combination ICS/LABAs (such as fluticasone furoate and vilanterol (BreoEllipta ^® ), fluticasone propionate/salmeterol (Advair ^® ), budesonide/formoterol (Symbicort ^® ), mometasone/formoterol (Dulera ^® ), ipratropium/albuterol sulfate (Duoneb ^® , Atrovent ^® ), albuterol/ipratropium (Combivent Respimat ^® )), SABAs (such as ipratropium (Atrovent ^® ) and albuterol sulfate (ProAir ^® , Proventil ^® )), and ICS (such as budesonide (Pulmicort ^® ) and fluticasone propionate (Flovent ^® ), beclomethasone dipropionate (QVAR ^® ).

Other agents that may be useful in combination therapies for treating COPD include SCH527123 (a CXCR2 antagonist), glycopyrrolate ((NVA237) Seebri ^® Breezhaler ^® ), glycopyrrolate and indacaterol maleate ((QVA149) Ultibro ^® Breezhaler ^® ), glycopyrrolate and formoterol fumarate (PT003), indacaterol maleate (QVA149), olodaterol (Striverdi ^® Respimat ^® ), tiotropium (Spiriva ^® )/olodaterol (Striverdi ^® Respimat ^® ), and aclidinium/formoterol inhaler.

In one embodiment of the invention, the at least one additional therapeutic agent is selected from oral corticosteroids, antithymocyte globulin, thalidomide, chlorambucil, calcium channel blockers, topical emollients, ACE inhibitors, serotonin reuptake inhibitors, endothelin-1 receptor inhibitors, antifibrotic agents, proton pump inhibitors or imatinib, ARG201 and tocilizumab. For example, in the treatment of systemic scleroderma, compounds that modulate STING, in particular compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with oral corticosteroids (such as prednisolone ( ^Delatsone® , Orapred, Millipred, Omnipred, Econopred, Flo-Pred), immunosuppressants (such as methotrexate ( ^Rhuematrex® , ^Trexall® ), cyclosporine ( ^Sandimmune® ), antithymocyte globulin ( ^Atgam® ), mycophenolate mofetil (CellCept®), cyclophosphamide ( ^Cytoxan® ), FK506 (tacrolimus), thalidomide ( ^Thalomid® ), chlorambucil (Leukeran®), azathioprine ( ^Imuran® , ^Azasan® )), calcium channel blockers (such as nifedipine (Procardia®, ^{Adalat®), tacrolimus (Tacrolimus®} ), thalidomide ( ^Thalomid® ), chlorambucil (Leukeran®), azathioprine (Imuran®, Azasan®), calcium channel blockers (such as nifedipine ( ^Procardia® , Adalat® ^{), tacrolimus (Tacrolimus ...} ) or nicardipine (Cardene ^® ), topical emollients (nitroglycerin ointment), ACE inhibitors (such as lisinopril (Zestril ^® , Prinivil ^® ), diltiazem (Cardizem ^® , Cardizem SR ^® , Cardizem CD ^® , Cardia ^® , Dilacor ^® , Tiazac ^® )), serotonin reuptake inhibitors (such as fluoxetine (Prozac ^® )), endothelin-1 receptor inhibitors (such as bosentan (Tracleer ^® ) or epoprostenol (Flolan ^® , Veletri ^® , Prostacyclin ^® )), antifibrotic agents (such as colchicine (Colcrys ^® ), para-aminobenzoic acid (PABA), dimethyl sulfoxide (DMSO), and D-penicillamine (Cuprimine ^® , Depen ^® ), interferon alpha and interferon gamma (INF-g), proton pump inhibitors (such as omeprazole (Prilosec ^® ), metoclopramide (Reglan ^® ), lansoprazole (Prevacid ^® ), esomeprazole (Nexium ^® ), pantoprazole (Protonix ^® ), rabeprazole (Aciphex ^® )) or imatinib (Gleevec ^® ) ARG201 (arGentis Pharmaceutical), belimumab (Benlysta ^® ), tocilizumab (Actema ^® ).

In one embodiment of the invention, at least one other therapeutic agent is ciliary neurotrophic growth factor or a gene transfer agent. For example, in the treatment of retinitis pigmentosa, a compound that modulates STING, particularly a compound of formula (I) or a pharmaceutically acceptable salt thereof, can be administered in combination with ciliary neurotrophic growth factor (NT-501-CNTF) or a gene transfer agent ^UshStat® .

In one embodiment of the invention, the at least one additional therapeutic agent is selected from a trivalent (IIV3) inactivated influenza vaccine, a quadrivalent (IIV4) inactivated influenza vaccine, a trivalent recombinant influenza vaccine, a quadrivalent live attenuated influenza vaccine, an antiviral agent or an inactivated influenza vaccine. For example, in the treatment of influenza, a compound that modulates STING, in particular a compound of formula (I ⁾ or a pharmaceutically acceptable salt thereof, can be administered in combination with a trivalent (IIV3) inactivated influenza vaccine (such as ^Afluria® , ^Fluarix® , ^Flucelvax® , ^FluLaval® , ^Fluvirin® , ^Fluzone® ), a quadrivalent (IIV4) inactivated influenza vaccine (such as Fluarix®, ^Flulaval® , ^Fluzone® ), a trivalent recombinant influenza vaccine (such as ^FluBlok® ), a quadrivalent live attenuated influenza vaccine (such as ^FluMist® ), an antiviral agent (such as oseltamivir ( ^Tamiflu® ), zanamivir ( ^Relenza® ), rimantadine ( ^Flumadine® ) or amantadine (Symmetrel® ⁾ ) or ^Fluad® , Fludase®, FluNhance® ^. , Preflucel or VaxiGrip ^® .

In the treatment of staphylococcal infections, compounds that modulate STING, in particular compounds of formula (I) or pharmaceutically acceptable salts thereof, can be administered in combination with antibiotics (such as β-lactam cephalosporins ( ^Duricef® , ^Kefzol® , Ancef®, ^{Biocef® ,} etc.), nafcillin ( ^Unipen® ), sulfonamides (sulfamethoxazole and trimethoprim (Bacrim®, ^Septra® , ⁾ , sulfasalazine ( ^Azulfidine® ), acetylsulfisoxazole (Gantrisin®), etc.) or vancomycin ( ^{Vancocin® )} ).

In one embodiment of the invention, the at least one additional therapeutic agent is selected from a topical immunomodulator or calcineurin inhibitor, a topical corticosteroid, an oral corticosteroid, interferon gamma, an antihistamine, or an antibiotic. For example, in the treatment of atopic dermatitis, compounds that modulate STING, particularly compounds of formula (I) or pharmaceutically acceptable salts thereof, may be administered in combination with a topical immunomodulator or calcineurin inhibitor such as pimecrolimus ( ^Elidel® ) or tacrolimus ointment ( ^Protopic® ), a topical corticosteroid such as hydrocortisone ( ^Synacort® , ^Westcort® ), betamethasone ( ^Diprolene® ), flurandrenolide (Cordan®), fluticasone (Cutivate®), triamcinolone (Kenalog® ⁾ , fluocinolone acetonide ( ^{Lidex® )} and clobetasol ( ^Temovate® ), an oral corticosteroid such as hydrocortisone ( ^Cortef® ), ^{methylprednisolone} ( ^Medrol® ) or prednisolone ( ^Pediapred® , Prelone® ^). ), immunosuppressants (such as cyclosporine (Neoral ^® ) or interferon gamma (Alferon N ^® , Infergen ^® , Intron A, Roferon-A ^® )), antihistamines (for itching, such as Atarax ^® , Vistaril ^® , Benadryl ^® ), antibiotics (such as the penicillin derivatives flucloxacillin (Floxapen ^® ) or dicloxacillin (Dynapen ^® ), erythromycin (Eryc ^® , T-Stat ^® , Erythra-Derm ^{® ,} etc.)), nonsteroidal immunosuppressants (such as azathioprine (Imuran ^® , Azasan ^® ), methotrexate (Rhuematrex ^® , Trexall ^® ), cyclosporine (Sandimmune ^® ) or mycophenolate mofetil (CellCept ^® )).

In the treatment of Parkinson's disease, the compounds of the invention may be administered in combination with L-dopamine based therapies (carbidopa (Lodosyn) - levodopa), dopamine agonists such as pramipexole (Mrapex), ropinirole (Requip), rotigotine (Neupro) and apomorphine (Apokyn), monoamine oxidase (MAO) B inhibitors such as selegiline (Eldepryl, Zelapar), rasagiline (Azilect) and safinamide (Xadago)), catechol O-methyltransferase (COMT) inhibitors entacapone (Comtan) and tolcapone (Tasmar), anticholinergics such as benztropine (Cogentin) or trihexylphenol oil and amantadine. The compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered in combination with a device implanted in a patient that delivers electrical pulses to the brain and reduces Parkinson's disease symptoms known as deep brain stimulation (DBS).

In the treatment of myocardial infarction, compounds of the invention may be administered in combination with anti-IL1β antibody therapy (eg, canakinumab).

The compounds of the invention may also be formulated as adjuvants with vaccines to modulate their activity. Such compositions may contain antibody (antibodies) or antibody fragment (antibody fragments) or antigenic components, including but not limited to proteins, DNA, live or dead bacteria and/or viruses or virus-like particles, and one or more components with adjuvant activity, including but not limited to aluminum salts, oil and water emulsions, heat shock proteins, lipid A preparations and derivatives, glycolipids, other TLR agonists such as CpG DNA or similar agents, cytokines such as GM-CSF or IL-12 or similar agents.

In a further aspect of the present invention, a vaccine adjuvant is provided, which comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof. A vaccine composition is further provided, which comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof and an antigen or an antigen composition.

A therapeutically "effective amount" is intended to mean an amount of a compound sufficient to effectively treat or prevent, as defined herein, when administered to a patient in need of such treatment. Thus, for example, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof is an amount of an agent of the invention sufficient to modulate the activity of STING such that a disease condition mediated by that activity is reduced, mitigated, or prevented when administered to a human in need thereof. The amount of a given compound corresponding to such an amount will vary depending on factors such as the specific compound (e.g., the efficacy ( _pIC50 ), potency ( _EC50 ), and biological half-life of the specific compound), the disease condition and its severity, the identity of the patient in need of treatment (e.g., age, size, and weight), but can still be determined routinely by one skilled in the art. Likewise, the duration of treatment with the compound and the time period of administration (time period between doses and timing of doses, e.g., before/with/after meals) will vary depending on the identity of the mammal to be treated (e.g., body weight), the specific compound and its properties (e.g., pharmacokinetic properties), the disease or condition and its severity, and the particular composition and method used, but can be determined by one skilled in the art.

"Treat" or "treatment" is intended to mean at least amelioration of a disease or condition in a patient. Methods of treatment for amelioration of a disease or condition include use of the compounds of the invention in any conventionally acceptable manner, e.g., for blocking, treating, or curing a STING-mediated disease or condition as described above. In one embodiment, "treat," "treating," or "treatment" with respect to cancer refers to amelioration of cancer, elimination or reduction of one or more symptoms of cancer, slowing or eliminating progression of cancer, and delaying recurrence of the condition in a patient or subject previously ill or diagnosed with the disease.

"Prevent," "prventing," or "preventment" refers to the prophylactic administration of a drug to reduce the likelihood of onset of a disease or its biological manifestation or to delay the onset of a disease or its biological manifestation. Prophylactic treatment is appropriate, for example, when a subject is considered to be at high risk for developing cancer, such as when the subject has a strong family history of cancer or when the subject has been exposed to a carcinogen.

The compounds of the present invention can be administered by any suitable route of administration, including systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration to the patient's lungs, whether by mouth or by inhalation through the nasal passages. Topical administration includes application to the skin.

In addition to the above-mentioned route of administration suitable for treating tumors, the pharmaceutical composition may be suitable for administration by intratumoral or paratumoral injection. It is expected that the compound of the present invention will be directly injected into a single solid tumor or near a tumor to trigger an immune response that can attack and destroy cancer cells in the whole body, substantially reducing and permanently eliminating tumors from patients with diseases in some cases. Activating the immune system in this way to kill tumors at distal sites is generally referred to as accompanying abscopal effects, and has been confirmed in animals with a variety of treatments ( van der Jeught, et al., Oncotarget, 2015, 6 (3), 1359-1381 ). A further advantage of local or intratumoral or paratumoral administration is the ability to achieve the same effect at a much lower dose, thereby minimizing or eliminating adverse events observable at much higher systemic doses (Marabelle, A., et al., Clinical Cancer Research, 2014, 20 (7), p1747-1756).

The compounds of the present invention may be administered via eye drops for the treatment of Sjögren's syndrome.

The compounds of the present invention can be administered once or according to a dosage regimen in which multiple doses are administered at different time intervals for a given period of time. For example, the dosage can be administered one, two, three or four times a day. The dosage can be administered until the desired therapeutic effect is achieved, or administered indefinitely to maintain the desired therapeutic effect. The appropriate dosage regimen of the compounds of the present invention depends on the pharmacokinetic properties of the compound, such as absorption, distribution and half-life, which can be determined by a technician. In addition, the appropriate dosage regimen of the compounds of the present invention (including the duration of administering such a regimen) depends on the severity of the disease or condition treated, the disease or condition treated, the age and physical condition of the patient treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic effect and similar factors within the technician's knowledge and profession. Such a technician should further understand that the appropriate dosage regimen can be adjusted according to the response of individual patients to the dosage regimen or because individual patients need to change over time. The total daily dose range is 1 mg to 2000 mg.

For use in therapy, the compounds of the invention will generally (but not necessarily) be formulated into a pharmaceutical composition prior to administration to a patient. Accordingly, the present invention also relates to a pharmaceutical composition comprising a compound of the invention and at least one pharmaceutically acceptable excipient.

The pharmaceutical composition of the present invention can be prepared and packaged in bulk form, wherein an effective amount of the compound of the present invention can be extracted and then given to the patient, such as with powder, syrup and solution for injection. Alternatively, the pharmaceutical composition of the present invention can be prepared and packaged into unit dosage form. For oral use, for example, one or more tablets or capsules can be administered. The dosage of the pharmaceutical composition contains at least a therapeutically effective amount of the compound of the present invention (i.e., a compound of formula (I) or a salt thereof, particularly a pharmaceutically acceptable salt).

As provided herein, unit dosage forms (pharmaceutical compositions) containing 1 mg to 1000 mg of a compound of the invention can be administered one, two, three, or four times daily to effectively treat a STING-mediated disease or condition.

The pharmaceutical compositions of the present invention generally contain one compound of the present invention. However, in certain embodiments, the pharmaceutical compositions of the present invention contain more than one compound of the present invention. In addition, the pharmaceutical compositions of the present invention may optionally further comprise one or more additional therapeutic agents (e.g., pharmaceutically active compounds).

As used herein, "pharmaceutically acceptable excipient" refers to a pharmaceutically acceptable substance, composition or vehicle that is involved in the form of administration or is compatible with the pharmaceutical composition. When mixed, each excipient must be compatible with the other ingredients of the pharmaceutical composition so that interactions that would substantially reduce the efficacy of the compounds of the invention when administered to a patient and interactions that would cause the pharmaceutical composition to become pharmaceutically unacceptable are avoided. In addition, the purity of each excipient must of course be high enough to make it pharmaceutically acceptable.

The compounds of the invention and one or more pharmaceutically acceptable excipients will generally be formulated into dosage forms suitable for administration to a patient via the desired route of administration. Conventional dosage forms include those that are: (1) suitable for oral administration, such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; (2) suitable for parenteral administration, such as sterile solutions, suspensions, and powders for reconstitution; (3) suitable for transdermal administration, such as transdermal patches; (4) suitable for rectal administration, such as suppositories; (5) suitable for inhalation, such as aerosols and solutions; and (6) suitable for topical administration, such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.

Suitable pharmaceutically acceptable excipients will vary according to the selected specific dosage form. In addition, suitable pharmaceutically acceptable excipients can be selected according to the specific functions they play in the composition. For example, certain pharmaceutically acceptable excipients can be selected according to their ability to promote the production of uniform dosage forms. Certain pharmaceutically acceptable excipients can be selected according to their ability to promote the production of stable dosage forms. Certain pharmaceutically acceptable excipients can be selected according to their ability to promote one or more compounds of the present invention to carry or transport to another organ or body part from one organ or body part once they are applied to the patient. Certain pharmaceutically acceptable excipients can be selected according to their ability to enhance patient compliance.

Suitable pharmaceutically acceptable excipients include excipients of the following types: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, cosolvents, suspending agents, emulsifiers, sweeteners, flavoring agents, taste masking agents, coloring agents, anti-caking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants and buffers. It will be appreciated by the skilled person that some pharmaceutically acceptable excipients may perform more than one function and may perform alternative functions, depending on how much the excipient is present in the formulation and what other ingredients are present in the formulation.

The skilled person has the knowledge and skill in the art to enable them to select suitable pharmaceutically acceptable excipients for use in the present invention in appropriate amounts. In addition, there are many resources available to the skilled person that describe pharmaceutically acceptable excipients and can be used to select suitable pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The pharmaceutical compositions of the present invention are prepared using techniques and methods known to those skilled in the art. Some methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

On the one hand, the present invention relates to solid oral dosage forms, such as tablets or capsules, which contain an effective amount of the compounds of the present invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g., corn starch, potato starch, and pregelatinized starch), cellulose and its derivatives (e.g., microcrystalline cellulose), calcium sulfate, and calcium hydrogen phosphate. Oral solid dosage forms may further include adhesives. Suitable adhesives include starch (e.g., corn starch, potato starch, and pregelatinized starch), gelatin, gum arabic, sodium alginate, alginic acid, tragacanth gum, guar gum, polyvidone, and cellulose and its derivatives (e.g., microcrystalline cellulose). Oral solid dosage forms may further include disintegrants. Suitable disintegrants include crospovidone, sodium starch glycolate, cross-linked carboxymethyl cellulose, alginic acid, and sodium carboxymethyl cellulose. Oral solid dosage forms may further include lubricants. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.

It will be appreciated that the compounds of the invention may also be formulated as adjuvants with vaccines to modulate their activity. Such compositions may contain antibody(ies) or antibody fragment(s) or antigenic components, including, but not limited to, proteins, DNA, live or dead bacteria and/or intact, inactivated or lysed viruses or virus-like particles, recombinant proteins or antigenic fragments thereof, optionally together with one or more other components having adjuvant activity, including, but not limited to, aluminum salts, oil and water emulsions, heat shock proteins, saponins, lipid A preparations and derivatives, glycolipids, liposomes, TLR agonists such as CpG DNA or similar agents, cytokines such as GM-CSF or IL-12 or similar agents.

Certain compounds of the invention may be potent immunomodulators and, therefore, should be handled with caution.

Example

The following examples illustrate the present invention. These examples are not intended to limit the scope of the present invention, but provide guidance for the technical staff on the preparation and use of the compounds, compositions and methods of the present invention. Although specific embodiments of the present invention have been described, the technical staff will appreciate that various changes and modifications may be made without departing from the spirit and scope of the present invention.

It will be appreciated that certain compounds of the invention may be potent immunomodulators and, therefore, should be handled with caution.

The reactions described herein are suitable for producing compounds of the invention having a variety of different substituent groups (e.g., R ¹ , R ² , etc.) as defined herein. It will be appreciated by the skilled artisan that if a particular substituent is incompatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. Suitable protecting groups and methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples thereof can be found in TW Greene 'Protective Groups in Organic Synthesis' (4th edition, J. Wiley and Sons, 2006). Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification.

Certain intermediate compounds described herein form yet further aspects of the invention.

General Synthesis Methods

The compounds of the present invention can be prepared using the synthetic procedures described in the following reaction schemes, which can be easily adjusted to prepare other compounds of the present invention by drawing on the knowledge of skilled organic chemists. The synthesis provided in these schemes is suitable for producing compounds of the present invention with various R groups using appropriate precursors, which, if necessary, can be suitably protected to achieve compatibility with the reactions outlined herein. If necessary, subsequent deprotection provides compounds of generally disclosed properties. Although the schemes are shown only with compounds of formula (I), they are illustrative methods that can be used to prepare the compounds of the present invention. Intermediates (compounds for preparing the compounds of the present invention) can also exist as salts.

Method 1

All variables are defined as in formula (I). Appropriately substituted halo-nitrophenyl compounds (1A) react with mono-protected diamines such as (1B) to provide N-protected nitroanilines. Removal of the amino protecting group yields amine (1D). Alternatively, amine (1D) can be directly obtained by reacting halo-nitrophenyl compounds (1A) with symmetrical diamines (1C). Amine (1D) can be reacted with halo-nitrophenyl compounds (1E) to obtain bis-nitro compounds (1F). In the case where (1A) is the same as (1E), bis-nitro compounds (1F) can be directly obtained by reacting diamine (1C) with excess halo-nitrophenyl compounds. Reduction of both nitro groups will provide tetraphenylamine (1G). Tetraphenylamine (1G) can be converted to (1H), an amidobenzimidazole dimer or macrocycle, by one of two methods: 1) treatment with cyanogen bromide to obtain a bis-aminobenzimidazole followed by coupling of the amide with a pyrazole acid such as (1K) or a linked-pyrazole diacid (1L); or 2) treatment with an isothiocyanate (1M) until dithiourea is completely formed, followed by addition of EDC (or other desulfurization reagent) and triethylamine (or other suitable base) and stirring until cyclodesulfurization is complete. Alkylation of the benzimidazole group of (1H) with an alkylating agent and a suitable base provides compounds of structure (1I). The site and extent of alkylation (i.e., mono-, di-, tri-alkylation) can generally be controlled by the choice of conditions. When suitable functional groups are present on (1I), deprotection or further functionalization of these groups will likely provide additional compounds (1J).

Solution 1

Method 2

All variables are defined as in formula (I). Appropriately substituted halo-nitrophenyl compounds (2A) react with mono-protected diamines such as (2B) to provide nitro-aniline (2C). Reduction of the nitro group under appropriate conditions yields diphenylamine (2D). Diphenylamine (2D) can be converted to amidobenzimidazole (2E) via one of two methods: 1) treatment with cyanogen bromide followed by coupling of the amide with a pyrazole acid such as (2M); or 2) treatment with an isothiocyanate (2N) until thiourea is fully formed, followed by addition of EDC (or other suitable desulfurization reagent) and triethylamine (or other suitable base) and stirring until desulfurization cyclization is complete. Removal of the amine protecting group yields amine (2F), which can react with halo-nitrophenyl compounds (2G) to yield nitroaniline (2H). Reduction of the nitro group will provide diphenylamine (2I). Diphenylamine (2I) can be converted to amidobenzimidazole (2J) via one of two methods: 1) treatment with cyanogen bromide followed by coupling of the amide with a pyrazole acid such as (2O); or 2) treatment with an isothiocyanate (2P) until thiourea is fully formed, followed by addition of EDC (or other suitable desulfurization reagent) and triethylamine (or other suitable base) and stirring until desulfurization and cyclization are complete. Alkylation of the benzimidazole group of (2J) with an alkylating agent and a suitable base provides compounds of structure (2K). The site and extent of alkylation (i.e., mono-, di-, tri-alkylation) can generally be controlled by the choice of conditions. When suitable functional groups are present on (2K), deprotection or further functionalization of these groups will likely provide additional compounds (2L).

Solution 2

Method 3

All variables are defined as in formula (I). Appropriately substituted halo-nitrophenyl compounds (3A) react with suitable dielectrophiles such as dibromide (3B) to provide nitro-aniline monobromide (3C). Treatment of (3C) with another appropriately substituted halo-nitrophenyl compound (3D) provides a linked dihalo-nitrophenyl compound (3E). Reaction of dihalo-nitrophenyl compounds (3E) with diamines (3F) containing linking groups provides dinitro macrocycles (3G). Reduction of both nitro groups will provide tetraphenylamine (3H). Tetraphenylamine (3H) can be converted to the macrocycle (3I) via one of two methods: 1) treatment with cyanogen bromide to provide a bis-aminobenzimidazole followed by coupling of the amide with a pyrazole acid such as (3L) or a linked-pyrazole diacid (3M); or 2) treatment with an isothiocyanate (3N) until dithiourea formation is complete, followed by addition of EDC (or other desulfurization reagent) and triethylamine (or other suitable base) and stirring until desulfurization and cyclization are complete. Alkylation of the benzimidazole group of (3I) with an alkylating agent and a suitable base provides compounds of structure (3J). The site and extent of alkylation (i.e., mono-, di-, tri-alkylation) can generally be controlled by the choice of conditions. When suitable functional groups are present on (3J), deprotection or further functionalization of these groups will likely provide additional compounds (3K).

Solution 3

Method 4

All variables are defined as in formula (I). Appropriately substituted halo-nitrophenyl compounds (4A) are reacted with mono-protected diamines such as (4B) to provide nitro-anilines (4C). Reduction of the nitro group under appropriate conditions will provide diphenylamines (4D). Diphenylamines (4D) can be converted to amide benzimidazoles (4E) via one of two methods: 1) treatment with cyanogen bromide followed by coupling of the amide with a pyrazole acid such as (4N); or 2) treatment with an isothiocyanate (4O) until thiourea is fully formed, followed by addition of EDC (or other suitable desulfurization reagent) and triethylamine (or other suitable base) and stirring until desulfurization cyclization is complete. Alkylation of the benzimidazole group of (4E) with an alkylating agent and a suitable base provides compounds of structure (4F). Deprotection/functionalization of the R group is performed if desired. Removal of the amide protecting group provides an amine (4G) which can react with a halo-nitrophenyl compound (4H) to provide a nitroaniline (4I). Reduction of the nitro group will provide the diphenylamine (4J). The diphenylamine (4J) can be converted to the amidobenzimidazole (4K) by one of two methods: 1) treatment with cyanogen bromide followed by coupling of the amide with a pyrazole acid such as (4P); or 2) treatment with an isothiocyanate (4Q) until the thiourea is fully formed, followed by addition of EDC (or other suitable desulfurization reagent) and triethylamine (or other suitable base) and stirring until desulfurization and cyclization are complete. Alkylation of the newly formed benzimidazole group of (4K) with an alkylating agent and an appropriate base provides compounds of structure (4L). The R group and/or linking group can be deprotected or functionalized (i.e., dihydroxylation of the linking group containing the olefin moiety) as desired to provide additional compounds (4M).

Solution 4

Method 5

All variables are defined as in formula (I). Appropriately substituted halo-nitrophenyl compounds (5A) react with mono-protected diamines such as (5B) to provide N-protected nitro-aniline. Removal of the amine protecting group provides amine (5D). Alternatively, amine (5D) can be directly obtained by reacting halo-nitrophenyl compounds (5A) with symmetrical diamines (5C). Amine (5D) can react with halo-nitrophenyl compounds (5E) to provide bis-nitro compounds (5F). In the case where (5A) is identical to (5E), bis-nitro compounds (5F) can be directly obtained by reacting diamines (5C) with excess halo-nitrophenyl compounds. If necessary, deprotection/functionalization of the R group (i.e., when the R ^A1 group is a halide, a C-C bond is formed). Reduction of the nitro group will provide tetraphenylamine (5H). If present and depending on the conditions used, other groups present in (5F) may also be reduced (i.e., olefins, aryl halides). Tetraphenylamine (5H) can be converted to (5I), an amidobenzimidazole dimer or macrocycle, by one of two methods: 1) treatment with cyanogen bromide to provide a bis-aminobenzimidazole followed by coupling of the amide with a pyrazole acid such as (5L) or a linked-pyrazole diacid (5M); or 2) treatment with an isothiocyanate (5N) until dithiourea formation is complete, followed by addition of EDC (or other desulfurization reagent) and triethylamine (or other suitable base) and stirring until desulfurization and cyclization are complete. Alkylation of the benzimidazole group of (5I) with an alkylating agent and a suitable base provides compounds of structure (5J). The site and extent of alkylation (i.e., mono-, di-, tri-alkylation) can generally be controlled by the choice of conditions. The R group and/or linking group can be deprotected or functionalized (i.e., dihydroxylation of the linking group containing the olefin moiety) as desired to provide additional compounds (5K).

Solution 5

The names of the intermediates and final compounds described herein were generated using the software naming program ChemDraw Pro 12.0.2.1076 Plug-In within Perkin Elmer E-Notebook or MarvinSketch 5.11.4_b82 (Chemaxon).

Those skilled in the art will appreciate that in some cases, these programs may name a structurally depicted compound as a tautomer or isomer of the compound. It will be understood that any reference to a named compound or a structurally depicted compound is intended to encompass all tautomers or isomers of such a compound and any mixtures of tautomers and/or isomers thereof.

The following abbreviations may be used in this manual:

abbreviation meaning AcOH Acetic acid aq. Watery BBr ₃ Boron tribromide BOC、tBOC tert-Butyloxycarbonyl brine Saturated sodium chloride aqueous solution Bu Butanol CDCl ₃ Deuterated chloroform CDI 1,1'-Carbonyldiimidazole _CH2Cl2 or _DCM Methylene chloride or dichloromethane CH ₃ CN or MeCN Acetonitrile CH ₃ NH ₂ Methylamine d sky DAST Diethylaminosulfur trifluoride DCE 1,2-Dichloroethane DCM Dichloromethane DIEA or DIPEA Diisopropylethylamine DMA dimethylacetamide DMAP 4-Dimethylaminopyridine DMF N,N -Dimethylformamide DMSO Dimethyl sulfoxide EDC 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide equiv equivalent Et Ethyl _Et3N or TEA Triethylamine _Et2O Diethyl ether EtOAc Ethyl acetate EtOH Ethanol FCC Flash column chromatography h、hr Hour HATU O -(7-Azabenzotriazol-1-yl)- N,N,N',N' -tetramethyluronium hexafluorophosphate HCl hydrochloric acid HOAt 1-Hydroxy-7-azabenzotriazole HOB Hydroxybenzotriazole HPLC High performance liquid chromatography ICl Iodine monochloride IPA Isopropyl alcohol i - _Pr2NEt N',N' -Diisopropylethylamine K ₂ CO ₃ Potassium carbonate KHMDS Potassium bis(trimethylsilyl)amide KO t -Bu Potassium tert-butoxide KOH Potassium hydroxide LCMS Liquid chromatography-mass spectrometry LiAlH ₄ Lithium aluminum hydride LiHDMS Lithium Hexamethyldisilazide LiOH Lithium hydroxide Me methyl MeOH or _CH3OH Methanol MgSO ₄ Magnesium sulfate min minute MS Mass spectrometry µw microwave NaBH ₄ Sodium borohydride _{Na2CO3 ​} Sodium carbonate NaHCO ₃ Sodium bicarbonate NaOH Sodium hydroxide _{Na2SO4 ​} Sodium sulfate NBS N-Bromosuccinimide _{N2H2 ​} Hydrazine NH ₄ Cl Ammonium chloride _NH4OH Ammonium Hydroxide NiCl ₂ •6H ₂ O Nickel(II) chloride hexahydrate NMO N-Methylmorpholine-N-oxide NMP N -Methyl-2-pyrrolidone NMR Nuclear Magnetic Resonance Pd/C Palladium on Carbon Ph Phenyl POCl ₃ Phosphorus oxychloride PSI Pounds per square inch RB Round bottom rm or rxn mixture Reaction mixture rt/RT Room temperature satd. Saturated sm Starting Materials TBAF Tetra-n-butylammonium fluoride TFA Trifluoroacetic acid THF Tetrahydrofuran TMEDA Tetramethylethylenediamine TMSI Trimethylsilyl iodide TMSN ₃ Trimethylsilyl azide T3P 2,4,6-Tripropyl-1,3,5,2,4,6-trioxatriphosphinane-2,4,6-trioxide t _R or Rf or Rt Retention time TxD p-Toluenesulfonic acid

Intermediate 1

(3-Bromopropoxy)(tert-butyl)dimethylsilane

3-bromopropane-1-ol (13.7 g, 99 mmol) was added to 1H-imidazole (13.4 g, 197 mmol) in DCM (100 mL), and then tert-butylchlorodimethylsilane (17.8 g, 118 mmol) was slowly added to DCM (20 ml). After 3 h at room temperature, the reaction was concentrated to ~100 mL and poured into EtOAc (800 mL), washed with 5% citric acid aqueous solution (2 x 200 mL) and brine. The organic layer was dried over MgSO ₄ , filtered and concentrated to obtain the title compound (10.0 g, 39.5 mmol, 40% yield). ¹ H NMR (400 MHz, chloroform- d ) δ ppm 3.78 (t, J =5.70 Hz, 2 H), 3.56 (t, J =6.46 Hz, 2 H), 2.07 (t, J =5.83 Hz, 2 H), 0.94 (s, 9 H), 0.11 (s, 6H).

Intermediate 2

4-Chloro-3-methoxy-5-nitrobenzamide

Methyl 4-chloro-3-methoxy-5-nitrobenzoate (1000 mg, 4.07 mmol) was stirred in NH ₄ OH (10 mL, 77 mmol) at room temperature for 24 h. Then, the reaction temperature was raised to 50 ^° C for 2 h. An additional 2 mL (~3.7 eq) of NH ₄ OH was added to the vessel. After stirring at 50 ° C for another 2 h (4 h total), the reaction was cooled to room temperature. The solid was filtered and rinsed with cold water. The solid was dried under house vacuum and lyophilized to provide 4-chloro-3-methoxy-5-nitrobenzamide (710 mg, 2.99 mmol, 73% yield) as a brown solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.31 (br. s., 1 H), 8.06 (d, J =1.77 Hz, 1 H), 7.88 (d, J =1.77Hz, 1 H), 7.81 (br. s., 1 H), 4.02 (s, 3 H). LCMS [M+H] ⁺ = 230.9.

Intermediate 3

4-Chloro-3-hydroxy-5-nitrobenzamide

4-Chloro-3-methoxy-5-nitrobenzamide (1 g, 4.34 mmol) was suspended in dry DCM (15 mL) and stirred at room temperature. BBr ₃ (17.4 mL, 1 M in DCM) was added dropwise to the reaction. A slurry was quickly formed, which was stirred overnight at room temperature under nitrogen. The reaction was poured into ice water (300 mL) and stirred vigorously for 30 min. The resulting suspension was filtered and the solid was dried to provide the title compound (610 mg, 2.82 mmol, 65% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 11.53 (br. s., 1 H), 8.17 (br. s., 1 H), 7.92 (s, 1 H), 7.72 (s, 1 H), 7.66 (br. s., 1 H). LCMS [M+H] ⁺ = 217.

Intermediate 4

3-(3-((tert-Butyldimethylsilyl)oxy)propoxy)-4-chloro-5-nitrobenzamide

(3-Bromopropoxy)(tert-butyl)dimethylsilane (7.3 g, 28.8 mmol) was dissolved in dry DMF (75 mL), 4-chloro-3-hydroxy-5-nitrobenzamide (4.8 g, 22.16 mmol) was added, followed by K ₂ CO ₃ (6.13 g, 44.3 mmol) and stirred at 100 °C under nitrogen for 2 h. The reaction was cooled to room temperature, poured into EtOAc (600 mL), washed with water (600 mL), brine, dried over MgSO ₄ , filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with 20-80% hexanes/EtOAc to give the title compound (7.43 g, 19.1 mmol, 86% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.29 (br. s., 1 H), 8.05 (d, J =1.71 Hz, 1H), 7.89 (d, J =1.71 Hz, 1 H), 7.77 (br. s., 1 H), 4.30 (t, J =5.99 Hz, 2 H) ,3.80 (t, J =5.99 Hz, 2 H), 1.98 (quint, J =5.99 Hz, 2 H), 0.80 - 0.90 (m, 9H), 0.02 (s, 6 H). LCMS [M + H] ⁺ = 389.

Intermediate 5

4-Chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide

A mixture of 4-chloro-3-hydroxy-5-nitrobenzamide (5000 mg, 23.09 mmol), 4-(3-chloropropyl)morpholine (4534 mg, 27.7 mmol), K2CO3 (4148 mg, 30.0 mmol) in DMF (30 mL) was stirred at 70°C overnight. The solvent was removed under vacuum to provide a crude solid product, which was purified by silica gel chromatography (12 g column, MeOH:DCM=1:10). The pure fractions were combined and the solvent was removed in vacuum to provide 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (3200 mg, 9.31 mmol, 40.3% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ )δ ppm 8.29 (s, 1 H), 8.04 (s, 1 H), 7.88 (d, J =1.2 Hz, 1 H), 7.77 (s, 1 H), 4.28 (t, J =6.2 Hz, 2 H), 3.62 - 3.52 (m, 4 H), 2.46 - 2.44 (m, 2 H), 2.37 (br. s., 4 H), 2.02 - 1.90 (m, 2 H). LCMS ( m/z ): 343.8 [M + H] ⁺ .

Intermediate 6

( E )-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide hydrochloride

Step 1: ( E )-tert-butyl (4-((4-carbamoyl-2-nitrophenyl)amino)but-2-en-1-yl)carbamate

A mixture of 4-fluoro-3-nitrobenzamide (10.0 g, 54.3 mmol), ( E )-tert-butyl (4-aminobut-2-en-1-yl) _carbamate (10.62 g, 57.0 mmol) and _K2CO3 (15.01 g, 109 mmol) in DMSO (200 mL) was stirred overnight at room temperature. The reaction was poured into water (2000 mL) and stirred for 30 min. The resulting solid was collected by filtration to give the title compound (18.3 g, 52.2 mmol, 96% yield). LCMS [2M+H] ⁺ = 700.5.

Step 2: ( E )-tert-butyl (4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)carbamate

To tert-butyl ( E )-(4-((4-carbamoyl-2-nitrophenyl)amino)but-2-ene-1-yl)carbamate (18.3 g, 52.2 mmol) in DMF (300 mL) was added stannous chloride dihydrate (58.9 g, 261 mmol). After stirring overnight at room temperature, the reaction was added dropwise to saturated NaHCO _aqueous solution (2000 mL) and extracted with EtOAc (5 x 500 mL). The combined organic layers were washed with brine (200 _mL ), dried over _Na2SO4 , filtered and concentrated to give the title compound (16.5 g, 51.5 mmol, 99% yield) as a yellow oil. LCMS [M-BOC+H] ⁺ = 221.1.

Step 3: ( E )-tert-butyl (4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

A mixture of ( E )-tert-butyl(4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)carbamate (16.5 g, 51.5 mmol) and cyanogen bromide (8.18 g, 77 mmol) in THF (200 mL) was heated to reflux overnight. The reaction was cooled to room temperature, diluted with saturated aqueous _NaHCO3 solution (500 mL), and extracted with EtOAc (5 x 300 mL). The combined organic layers were washed with brine, dried over _Na2SO4 , filtered and concentrated. The residue was purified over silica gel eluting with 50: ₁ to 20:1 DCM in MeOH (+ 3% _NH4OH ) to give the title compound (13.7 g, 39.7 mmol, 77% yield) as an off-white solid. LCMS [M+H] ⁺ = 346.1.

Step 4: ( E )-tert-butyl (4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

To 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (9.17 g, 59.5 mmol) in DCM (500 mL) was added EDC (20.53 g, 107 mmol) and HOBt (18.22 g, 119 mmol) at 0°C. After 15 min, a mixture of ( E )-tert-butyl(4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (13.7 g, 39.7 mmol) in DMF (50 mL) was added, followed by TEA (27.6 mL, 198 mmol). The reaction was warmed to room temperature, stirred overnight and concentrated. The residue was diluted with water (500 mL) and extracted with EtOAc (3 X 300 mL), and the combined organic phases were washed with brine, dried _{over Na2SO4} , filtered and concentrated. The residue was purified on silica gel eluting with 50:1 to 20:1 DCM:MeOH to provide the crude product, which was washed with DCM (300 mL) and collected by filtration to give the title compound as an off-white solid (14.0 g, 29.1 mmol, 73% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δppm 12.84 (s, 1 H), 8.00 - 7.97 (m, 2 H), 7.80 - 7.78 (m, 1 H), 7.49 (d, J =8.4 Hz, 1 H), 7.34 (s, 1 H), 6.95 (t, J =5.5 Hz, 1 H), 6.66 (s, 1 H), 5.73 -5.65 (m, 2 H), 4.83 (d, J =4.3 Hz, 2 H), 4.62 (q, J =7.0 Hz, 2 H), 3.52 (s, 2H), 2.18 (s, 3 H), 1.38 - 1.33 (m, 12 H); LCMS [M + H] ⁺ = 482.0.

Step 5: ( E )-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide hydrochloride

To a suspension of tert-butyl ( E )-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (3.00 g, 6.23 mmol) in dioxane (60 mL) was added 4 N HCl in dioxane (15.6 mL, 62.3 mmol) followed by MeOH (15 mL) to dissolve some of the remaining solid. After 30 min at room temperature, the reaction mixture became cloudy and was stirred for approximately 3 days. The resulting solid was collected by filtration and washed with DCM to obtain the title compound as a white solid (2.0 g, 4.8 mmol, 77% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.97 - 8.09 (br. s., 1H), 7.82 (d, J =8.11 Hz, 1 H), 7.50 (d, J =8.11 Hz, 1 H), 7.38 (br. s., 1 H), 6.70 (s, 1 H), 5. 97 - 6.08 (m, 1 H), 5.68 - 5.80 (m, 1 H), 4.91 (d, J =4.31Hz, 2 H), 4.60 (q, J =6.67 Hz, 2 H), 3.42 (br. s., 2 H), 2.18 (s, 3 H), 1.36(t, J =6.97 Hz , 3 H); LCMS [M+H] ⁺ = 382.2.

Intermediate 7

( E )-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide

Step 1: ( E )-1-(4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

A microwave tube containing ( E )-1-(4- _aminobut -2-en-1- _yl )-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide hydrochloride (517 mg, 1.24 mmol) in DMSO (10 mL) was treated with TEA (0.28 mL, 2.0 mmol) followed by K 2 CO 3 (274 mg, 1.98 mmol) and 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-chloro-5-nitrobenzamide (385 mg, 0.990 mmol). The reaction was heated to 75° C. After 7 h, the mixture was concentrated and the residue was purified over silica gel eluting with 10-90% EtOAc (to remove impurities) and then 0-10% MeOH in DCM to give the title compound as an orange solid (200 mg, 0.273 mmol, 28% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.16 (d, J =1.52 Hz, 1 H), 7.94 - 8.08 (m, 3 H), 7.74 (d, J =8.11 Hz, 2 H), 7.50 (s, 1 H), 7.31 - 7.43 (m, 3 H), 6.62 (s, 1 H), 5.74 - 5.81 (m, 2 H), 4.80 (br. s., 2 H), 4.59 (d, J =6.84 Hz, 2 H), 4.13 (br. s., 2 H), 4.01 (t, J =6.08 Hz, 2 H), 3.63 (t, J =5.96Hz, 2 H ), 2.16 (s, 3 H), 1.76 - 1.88 (m, 2 H), 1.33 (t, J =7.10 Hz, 3 H), 0.74- 0.82 (m, 9 H), -0.06 (s, 6 H); LCMS [M + H] ⁺ = 734.6.

Step 2: ( E )-1-(4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

(E)-1-(4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (1 g, 1.363 mmol) was suspended in MeOH (20 mL), ammonium hydroxide (4.62 mL, 34.1 mmol) was added and stirred at room temperature for 5 min. Then sodium sulfoxylate (1.675 g, 8.18 mmol) in water (5 mL) was added. After 60 min, EtOAc (300 ml) was added and the mixture was extracted with water (50 ml x 3). The organic layer was separated, dried over Na ₂ SO ₄ , and concentrated under vacuum to afford the title compound (710 mg, 1.009 mmol, 74.0% yield) as a light yellow solid, which was used without further purification. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.80 (br. s., 1H), 8.00 (s, 1 H), 7.97 (br. s., 1 H), 7.75 (dd, J =8.49, 1.14 Hz, 1 H), 7.63(br. s., 1 H), 7.28 - 7.41 (m, 2 H), 7.00 (br. s., 1 H), 6.84 (d, J =1.52 Hz, 1 H), 6.74 (d, J =1.52 Hz, 1 H), 6.65 (s, 1 H), 5.79 - 5.96 (m, 1 H), 5.64 -5.78 (m, 1 H), 4.81 (d, J =4.82 Hz, 2 H), 4.68 (br. s., 2 H), 4.61 (d, J =7.10Hz, 2 H), 3.92 (t, J =5.83 Hz, 2 H), 3.84 (br. s., 1 H), 3.63 (t, J =6.08 Hz, 2H), 3.57 (br. s. , 2 H), 2.17 (s, 3 H), 1.70 - 1.82 (m, 2 H), 1.34 (t, J =7.10Hz, 3 H), 0.68 - 0.83 (m, 9 H), -0.06 (s, 6 H); LCMS [M + H] ⁺ = 704.3.

Step 3: ( E )-2-amino-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-1H-benzo[d]imidazole-5-carboxamide

At room temperature, ( E )-1-(4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-amino

To a solution of 2-(1-ethyl-3-methyl-1H-pyrazole-5-formamido)-1H-benzo[d]imidazole-5-carboxamide (120 mg, 0.170 mmol) in MeOH (5 mL) was added cyanogen bromide (36 mg, 0.34 mmol). After 2 h, the reaction was concentrated and EtOAc (10 mL) was added. After stirring for 30 min, the solid was isolated by filtration and washed with EtOAc to obtain the title compound as a light brown solid (120 mg, 0.165 mmol, 97% yield), which was used without further purification. ¹ H NMR (400 MHz, MeOH- d ₄ ) δ ppm 8.00 (d, J =1.27 Hz, 1 H), 7.81 (dd, J =8.36, 1.77 Hz, 1 H), 7.49 (d, J =1.27 Hz, 1 H), 7.39 - 7.45 (m, 1 H), 7.36 (d, J =1.27 Hz, 1 H), 6.61 (s, 1 H), 5.82 - 5.99 (m, 2 H), 4.96 - 5.01 (m, 2 H), 4.56 - 4.65 (m, 2 H), 4.12 (t, J =6.21 Hz, 2H), 3.62 - 3.75 (m, 2 H), 2.18 - 2.29 (m, 3 H), 1.79 (t, J =6.21 Hz, 2 H), 1.24 - 1.54 (m, 5 H), 0.84 - 0.98 (m, 9 H), -0.01 - 0.11 (m, 6 H); LCMS [M +H] ⁺ = 729.5.

Step 4: ( E )-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (33 mg, 0.21 mmol) in DMF (3 mL) was added HATU (75 mg, 0.20 mmol) and HOBt (12.6 mg, 0.082 mmol). After stirring at room temperature for 10 min, triethylamine (0.09 mL, 0.66 mmol) was added, followed by ( E )-2-amino-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-1H-benzo[d]imidazole-5-carboxamide (120 mg, 0.165 mmol) and the reaction was continued at room temperature. After 3 days, by dropwise addition of water, solid precipitated out from the reaction. Solid was separated by filtration and washed with water. Then, the solid was purified by eluting with 0-20% MeOH in DCM through silica gel (12g HP Gold column). Combine required fractions and concentrate to obtain the title compound (29 mg, 0.034 mmol, 20% yield) as off-white solid. ¹ HNMR (400 MHz, THF- d ₄ ) δ ppm 12.53 (br. s., 2 H), 8.00 (d, J =1.01 Hz, 1 H), 7.61 (d, J =1.01 Hz, 1 H), 7.53 (dd, J =8.36, 1.52 Hz, 1 H), 7.36 (d, J = 6.84Hz, 2 H), 7.29 (d, J =1.01 Hz, 1 H), 7.12 (d, J =8.36 Hz, 1 H), 6.83 (br. s., 2H), 6.66 (d, J =2.28 Hz, 2 H), 6.06 (dt, J =15.46, 5.58 Hz, 1 H), 5.87 (dt, J =15.46, 5.83 Hz, 1 H), 5.09 (d, J =5.32 Hz, 2 H), 4.89 (d, J =5.58 Hz, 2 H),4.59 - 4.72 (m, 4 H), 3.97 (t, J =6.21 Hz, 2 H), 3.69 (t, J =5.96 Hz, 2 H),2.20 (s, 6 H), 1.73 - 1.78 (m, 2 H), 1.40 (td, J =7.03, 1.14 Hz, 6 H), 0.82 -0.94 (m, 9 H), -0.03 - 0.09 (m, 6 H); LCMS [M/2+H] ⁺ = 433.6。

Intermediate 8

1-Ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate

To a 1 L round bottom flask was added 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (25 g, 162 mmol) and DCM (500 mL). To this heterogeneous solution was added DMF (0.1 mL, 1.291 mmol) followed by the slow addition of oxalyl chloride (15.61 mL, 178 mmol). After stirring at room temperature for 1 h, the volatiles were removed under vacuum and the crude material was co-evaporated twice with dichloromethane (100 mL each). Assume 100% yield and use the crude product directly (1-ethyl-3-methyl-1H-pyrazole-5-carbonyl chloride (28.0 g, 162 mmol, 100% yield)).

To a dry 1L round bottom flask was added KSCN (18.92 g, 195 mmol) and acetone (463 mL). This clear homogeneous solution was cooled to 0°C. After stirring at 0°C for 5 min, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl chloride (28 g, 162 mmol) was added as a solution in acetone (25 mL). Once the addition was complete, the reaction was allowed to stir at 0°C. After 1 min, additional KSCN (~2 g) was added and the reaction was stirred for an additional 20 min. At this point, hexanes (200 mL) were added to the reaction mixture and the crude heterogeneous solution was concentrated to one third of the volume in vacuo. The process of hexanes addition and concentration was repeated twice (300 mL of hexanes each time). After the last concentration, hexanes (200 mL) were added and the solids were removed by filtration, rinsing with hexanes (100 mL). The resulting clear light yellow filtrate was concentrated and purified by chromatography (330 g Gold silica column; eluted with 0-20% EtOAc/hexanes). The desired product eluted at ~7% EtOAc/hexanes. The desired fractions were combined and concentrated to give 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (27.5 g, 139 mmol, 86% yield) as a clear colorless liquid. ¹ H NMR (400 MHz, CHLOROFORM- d ) δ ppm 6.77 (s, 1 H), 4.54 (q, J =7.10 Hz, 2 H), 2.34 (s, 3 H), 1.44 (t, J =7.22 Hz, 3 H); LCMS [M + H] ⁺ = 196.1. The acyl isothiocyanate product degraded over time, so a ~0.4 M 1,4-dioxane solution was prepared and frozen to avoid/slow decomposition. This solution was thawed and used directly in subsequent reactions.

Intermediate 9

tert-Butyl (E)-(4-((4-Carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)carbamate

To a suspension of 4-chloro-3-methoxy-5-nitrobenzamide (1.50 g, 6.50 mmol) in EtOH (25 mL) was added ( E )-tert-butyl (4-aminobut-2-en-1-yl)carbamate (1.454 g, 7.81 mmol) and DIEA (3.4 mL, 20 mmol). The reaction was stirred at 120 °C in a sealed tube and allowed to cool to room temperature. The resulting orange solid was collected by filtration and washed with EtOH to give the title compound (2.10 g, 5.52 mmol, 85% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.19 (d, J=1.77 Hz, 1 H) 8.03 (br. s., 1 H) 7.76 (t,J=6.08 Hz, 1 H) 7.55 (d, J=1.52 Hz, 1 H) 7.34 (br. s., 1 H) 6.9 5 (t, J=5.45Hz, 1 H) 5.53 (br. s., 2 H) 4.09 (br. s., 2 H) 3.88 (s, 3 H) 3.48 (br. s., 2H) 1.35 (s, 9 H). LCMS ( m/z ): 325.1 [M- t -Bu + H] ⁺ .

[00136] Alternative route to prepare compounds described herein: To a 3-neck 5-liter flask was added 4-chloro-3-methoxy-5-nitrobenzamide (451.10 g, 1956 mmol), n-butanol (2174 mL), N-ethyl-N-isopropylpropan-2-amine (854 mL, 4890 mmol), and (E)-tert-butyl (4-aminobut-2-en-1-yl)carbamate (403.1 g, 2164 mmol). A condenser and a septum containing a temperature probe were attached to the flask. The reaction flask was stirred with an overhead stirrer (300 rpm) and heated to 110°C using a heating mantle attached to a thermostat. After 6 h, the heterogeneous mixture turned dark red and homogeneous. The reaction was stirred at 110°C for 24 h. The mixture was cooled to room temperature. Isopropanol (1200 mL) was added. The solid was filtered on a Buchner funnel. The organic filter cake was rinsed twice with isopropanol (1200 mL each time). The solid was air dried overnight (~14 h). (E)-(4-((4-carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-ene-1-yl)carbamic acid tert-butyl ester (532 g, 1343 mmol, 68.6% yield) was obtained as an orange solid. By 1H NMR, HPLC tracking and LCMS, the minimum purity of this solid was estimated to be 96%. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.19 (d, J =1.77Hz, 1 H) 8.03 (br. s., 1 H) 7.76 (t, J =5.96 Hz, 1 H) 7.56 (d, J =1.77 Hz, 1 H)7.35 (br. s., 1 H) 6.96 ( t, J =5.58 Hz, 1 H) 5.53 (br. s., 2 H) 4.09 (br. s., 2 H) 3.88 (s, 3 H) 3.48 (br. s., 2 H) 1.09 - 1.54 (m, 9 H). LCMS ( m/z ): 381.2[M + H] ⁺ .

Intermediate 10

( E )-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzamide, hydrochloride

To a suspension of tert-butyl (E)-(4-((4-carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)carbamate (20 g, 47.3 mmol) in MeOH (50 mL) was slowly added 4M HCl in dioxane (100 mL, 400 mmol). The reaction mixture was stirred at room temperature for 1 h, then the resulting solid was isolated by filtration, washed with diethyl ether (3 x 100 mL), and dried under high vacuum to afford the title compound (13.90 g, 43.9 mmol, 93% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.22 (d, J =2.03 Hz, 1 H), 7.76 - 8.16 (br. m., 5 H), 7.60 (d, J =2.03 Hz, 1 H), 7.37 (br. s., 1 H), 5.87 (dt, J =15.52 , 5.80 Hz, 1 H), 5.62 (dt, J =15.65, 6.37 Hz, 1 H), 4.18 (d, J =5.32 Hz, 2H), 3.90 (s, 3 H), 3.40 (t, J =5.70 Hz, 2 H). LCMS ( m/z ): 281.1 [M + H] ⁺ .

Intermediate 11

(E)-1-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 3 hydrochloride

Step 1: tert-Butyl ((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

To tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-ene-1-yl)carbamate (530 mg, 1.036 mmol) in DMF (5 mL) was added cesium carbonate (675 mg, 2.072 mmol) and iodomethane (0.097 mL, 1.554 mmol) at room temperature. The reaction was stirred at room temperature. After 2 h, the reaction was diluted with 100 mL EtOAc, washed with 2 x 100 mL water and 3 x 100 mL brine. The organic layer was collected and concentrated under vacuum to give the title compound (630 mg, 1.04 mmol, 100% yield) as a yellow solid. LCMS m/z = 526 [M + H] ⁺ .

Step 2: (E)-1-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 3-hydrochloride

To a solution of tert-butyl ((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (600 mg, 1.142 mmol) in MeOH (5 mL) was added 4 M hydrochloric acid in dioxane (2.85 mL, 11.42 mmol) and the reaction was stirred at room temperature. After 3 h, the volatiles were removed under vacuum to give the title compound as an orange solid (650 mg, 1.04 mmol, 92% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 1.33(t, J =7.10 Hz, 3 H) 2.20 (s, 3 H) 3.32 - 3.42 (m, 3 H) 3.66 (br. s., 3 H)4.03 (s, 3 H) 4.54 (q, J =7.10 Hz, 2 H) 5.03 (br. s., 2 H) 5.60 - 5.71 (m, 1H) 5.97 (dt, J =15.59, 5.89 Hz, 1 H) 6.79 (br. s., 1 H) 7.52 - 7.61 (m, 2 H)7.90 (br. s., 1 H) 8.05 (br. s., 3 H) 8.22 (br. s., 1 H). LCMS m/z = 426 [M +H] ⁺ .

Intermediate 12

( E )-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-5-carboxamide

Step 1: ( E )-4-((4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzamide

(E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzamide, hydrochloride (1.7 g, 5.37 mmol), 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (1.65 g, 4.81 mmol), isopropanol (15 mL), and DIPEA (2.94 mL, 16.85 mmol) were separated into two 24-mL vials. The vials were capped and heated at 120 °C for 42 h. The resulting solid was isolated by filtration, rinsing with isopropanol (2 x 3 mL) to afford ( E )-4-((4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzamide (1.95 g, 2.79 mmol, 51.9% yield) as a brick red solid. LCMS ( m/z ): 588.2 [M+H] ⁺ .

Step 2: ( E )-3-amino-4-((4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzamide

To ( E )-4-((4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzamide (4.6 g, 6.65 mmol) in MeOH (83.0 mL) was added sodium sulfoxylate (19.08 g, 93.0 mmol) in water (70 mL) at room temperature. After 15 min, solid sodium bicarbonate (24 g) was added. After 10 min, the reaction was filtered and the solid was rinsed with MeOH (4 x 20 mL). The combined filtrates were concentrated onto celite and purified by dry-loading onto silica gel (80 g Gold column), eluting with 2-40% (10:1 MeOH:aq. NH ₄ OH) in DCM to give the title compound (1.81 g, 3.26 mmol, 49% yield) as a dark yellow film. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.64 (br. s., 2 H), 6.99 (br. s., 2 H), 6.85 (dd, J =5.07, 1.77 Hz, 2 H), 6.78 (dd, J =4.31, 1.77 Hz, 2H), 5.63 - 5. 72 (m, 2 H), 4.66 (d, J =8.11 Hz, 4 H), 3.96 (t, J =6.21 Hz, 2 H), 3.74 (s, 3 H), 3.51 - 3.60 (m, 6 H), 3.17 (br. s., 4 H), 2.43 (t, J =7.10 Hz, 2 H), 2.35 (br. s., 4 H), 1.87 (t, J =6.72 Hz, 2 H); LCMS ( m/z ): 528.4 [M + H] ⁺ .

Step 3: ( E )-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-5-carboxamide

To ( E )-3-amino-4-((4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)-but-2-en-1-yl)amino)-5-methoxybenzamide (368 mg, 0.697 mmol) in DMF (6.97 mL) at 0° C. was added 0.4 M 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate in dioxane (2.0 mL, 0.80 mmol). After ~10 min, another portion of 0.4 M 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate in dioxane (0.5 mL, 0.20 mmol) was added and after ~15 min, the last portion (0.5 mL, 0.20 mmol) was added. After 35 min total reaction time, EDC (334 mg, 1.74 mmol) was added followed by triethylamine (0.486 mL, 3.49 mmol). The mixture was allowed to warm to room temperature and stirred overnight (~14 hours). The reaction was quenched with 3:1 water: saturated aqueous NH ₄ Cl solution (40 mL) and extracted with 3:1 chloroform: EtOH (2x40 mL). The combined organic phases were washed with water (20 mL), dried over magnesium sulfate and concentrated. The resulting residue was purified by silica gel chromatography (40 g column, 2-40% gradient of [10:1 MeOH: aqueous NH ₄ OH]/DCM) to provide the title compound (361 mg, 0.425 mmol, 60.9% yield) as a peach solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 1.20-1.35 (m, 6 H), 1.55 - 1.73 (m, 2 H), 2.02 - 2.31 (m, 12 H), 3.46 (t, J =4.44 Hz, 4 H), 3.70 (s, 3 H), 3.93 (t, J =5.96 Hz, 2 H), 4.40 - 4.68 (m, 4 H), 4.80 - 5.00 (m, 4H), 5.69 - 6.00 (m, 2 H), 6.41 - 6.74 (m, 2 H), 7.13 - 7.51 (m, 4 H), 7.56 -7.76 (m, 2 H), 7.99 (d, J =3.55 Hz, 2 H), 12.85 (br. s., 2 H). LCMS ( m/z ):851.5 [M + H] ⁺ .

Intermediate 13

2,2,3,3-Tetrafluorobutane-1,4-diamine

Step 1: 2,2,3,3-tetrafluorobutane-1,4-diyl bis(4-methylbenzenesulfonate)

4-Methylbenzene-1-sulfonyl chloride (29.4 g, 154 mmol) was added to 2,2,3,3-tetrafluorobutane-1,4-diol (10.0 g, 61.7 mmol) in pyridine (150 mL) at 0°C over 5 min, and the reaction was then heated to 55°C. After 1 day, the reaction was quenched with ice water, and the resulting solid was collected by filtration, dissolved in DCM (200 mL) and washed with 5% H ₂ SO ₄ aqueous solution (100 mL x 3). The organic layer was dried over Na ₂ SO ₄ and concentrated to give the title compound (27.3 g, 58.0 mmol, 94% yield) as a white solid. LCMS [M + H] ⁺ = 470.9.

Step 2: 1,4-Diazido-2,2,3,3-tetrafluorobutane

Bis(4-methylbenzenesulfonic acid) 2,2,3,3-tetrafluorobutane-1,4-diyl ester (10.0 g, 21.3 mmol) and sodium azide (5.53 g, 85.0 mmol) in DMF (40 mL) were stirred at 110 ° C overnight. The reaction was quenched with NaClO (aq) and extracted with DCM (5 mL x 3). The combined organic layer was washed with water (10 mL), dried over Na ₂ SO ₄ and concentrated to give the title compound (3.5 g, 16.5 mmol, 78% yield). LCMS [M + H] ⁺ = 213.1.

Step 3: 2,2,3,3-Tetrafluorobutane-1,4-diamine

To a solution of 1,4-diazide-2,2,3,3-tetrafluorobutane (36.0 g, 170 mmol) in MeOH (350 mL) was added 10% palladium on carbon (18.1 g, 17.0 mmol). The reaction mixture was stirred at 40 °C under hydrogen (4 atm) for 16 h. The mixture was filtered through a pad of celite, washed with MeOH and the filtrate was concentrated in vacuo to give the title compound (22.0 g, 124 mmol, 73% yield). ¹ H NMR (400 MHz, CHLOROFORM- d ) δ ppm 3.12 - 3.37 (m, 4H), 1.43 (br. s., 4 H).

Intermediates 14A and 14B

7-(3-((tert-Butyldimethylsilyl)oxy)propoxy)-1-(5-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide

Step 1: 2,5-Diazidohexane

To a 500-mL round-bottom flask was added 2,5-dibromohexane (10 g, 41.0 mmol) and DMF (100 mL). To this homogeneous solution was added sodium azide (10.66 g, 164 mmol). The heterogeneous reaction mixture was stirred at 80 °C for 1 h. The mixture was cooled to room temperature and water (100 mL) was added. The aqueous phase was extracted with diethyl ether (3 x 100 mL). The combined organic phases were washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated under vacuum. 2,5-diazidehexane (8.54 g, 33.5 mmol, 83% yield, 66% purity) was obtained as a yellow oil. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm1.30 (d, J=6.59 Hz, 6 H), 1.40 - 1.76 (m, 4 H), 3.35 - 3.68 (m, 2 H).

Step 2: Hexane-2,5-diamine

2,5-diazidohexane (8.54 g, 50.8 mmol) was dissolved in MeOH (300 mL). The solution was hydrogenated in a single pass through a ThalesNano H- ^Cube® system (35°C, 25 bar hydrogen pressure, 2 mL/min flow rate). The solution was then concentrated and the crude product was used in the next reaction. Hexane-2,5-diamine (6.04 g, 49.4 mmol, 97% yield) was obtained as a colorless oil. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 1.09 (dd, J =6.21, 1.65Hz, 6 H), 1.21 - 1.62 (m, 8 H), 2.78 - 3.02 (m, 2 H).

Step 3: 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-((5-((4-carbamoyl-2-nitrophenyl)amino)hexan-2-yl)amino)-5-nitrobenzamide

Into a 40-mL vial was placed 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-chloro-5-nitrobenzamide (Intermediate Compound 4) (1.255 g, 3.23 mmol), isopropanol (8 mL), and DIPEA (1.879 mL, 10.76 mmol). To this heterogeneous mixture was added hexane-2,5-diamine (500 mg, 4.30 mmol) as a solution in isopropanol (2 mL). The vial was capped and heated to 110 °C overnight (~14 h). The solution was cooled to room temperature. 4-Fluoro-3-nitrobenzamide (0.594 g, 3.23 mmol) was added followed by DIPEA (1.879 mL, 10.76 mmol). The reaction was heated to 110 °C again for 2 h. A solid formed after cooling to room temperature. The solid was collected on the filter and rinsed twice with isopropanol (2 mL each time). The crude solid was purified by silica gel chromatography (ISCO equipment, 80 g _SiO2 cartridge, 2-20% gradient of MeOH/DCM). The corresponding fractions were combined and concentrated. 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-((5-((4-carbamoyl-2-nitrophenyl)amino)hexane-2-yl)amino)-5-nitrobenzamide (300 mg, 0.450 mmol, 10.47% yield) was obtained as an orange glass film (mixture of diastereomers). LCMS ( m/z ): 633.5 [M + H] ⁺ .

Step 4: 3-amino-4-((5-((2-amino-4-carbamoylphenyl)amino)hexan-2-yl)amino)-5-(3-((tert-butyldimethylsilyl)oxy)propoxy)benzamide

To a 125-mL Erlenmeyer flask was added 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-((5-((4-carbamoyl-2-nitrophenyl)amino)hexan-2-yl)amino)-5-nitrobenzamide (386 mg, 0.610 mmol) and MeOH (40 mL). The solution was hydrogenated using a ThalesNano H- ^Cube® system (5% Pd/C cartridge, 30° C., 10 bar hydrogen pressure, 1.5 mL/min flow rate). After two cycles, the reduction was complete. The solution was concentrated to afford 3-amino-4-((5-((2-amino-4-carbamoylphenyl)amino)hexan-2-yl)amino)-5-(3-((tert-butyldimethylsilyl)oxy)propoxy)benzamide (352 mg, 0.602 mmol, 99% yield). LCMS ( m/z ): 573.5[M + H] ⁺ .

Step 5: 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To a 100-mL round-bottom flask was added 3-amino-4-((5-((2-amino-4-carbamoylphenyl)amino)hexan-2-yl)amino)-5-(3-((tert-butyldimethylsilyl)oxy)propoxy)benzamide (352 mg, 0.614 mmol) and DMF (6.1 mL). A solution of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (Intermediate Compound 8) (~0.4 M in dioxane, 2.75 mL, 1.100 mmol) was added at 0 °C and the mixture was stirred for 15 min. Then, EDC (295 mg, 1.536 mmol) was added followed by triethylamine (0.428 mL, 3.07 mmol). The reaction was stirred at room temperature overnight (~14 h). The reaction was distributed between 50 mL of ethyl acetate and 50 mL of a 1:1 mixture of saturated aqueous ammonium chloride and water. The layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 25 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated under vacuum. Purification by reverse phase preparative chromatography (biphasic ISCO system, GeminiC18, 5 um, 50 x 30 mm column; 40-70% gradient of MeCN/water (containing NH ₄ OH modifier)) enabled separation and characterization of the first eluting diastereomer and the second eluting diastereomer. Each diastereomer was expected to be a racemate (i.e., an enantiomeric pair). The fractions containing the first eluting diastereomer and the second eluting diastereomer were combined and dried to provide intermediates XA and XB as white solids, respectively.

Intermediate 14A (first eluting diastereomer)

Racemic 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (200 mg, 0.223 mmol, 36.4% yield).

LCMS ( m/z ): 895.6 [M + H] ⁺ ; 1.37 min retention time (Acquity UPLC CSH C18, 1.7 um, 50 mm x 2.1 mm column; 40°C; 3-95% gradient over 1.5 min, MeCN/10 mM ammonium bicarbonate in water adjusted to pH 10 with 25% ammonium hydroxide solution).

¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 7.98 (br. s., 0.54), 7.91 (s, 0.51), 7.80 (t, J =7.22 Hz, 1.06), 7.45 - 7.65 (m, 2.01 ), 7.40 (s, 0.52), 7.35 (s,0.54), 6.34 - 6.79 (m, 2.09), 5.44 (br. s., 1.20), 4.65 (m, 4.10), 4.28 (m,1.56), 3.99 (br. s., 0.63), 3.79 (m, 1.06), 3.70 (br. s., 0.65), 3.61 (br.s., 0.58), 2.98 (br.s., 2.98), 2.35 - 2.65 (m, 1.40), 2.28 (s, 1.49), 2.23(m, 3.16), 2.17 (br.s., 1.47 ), 1.99 (br. s., 0.98), 1.81 (br. s., 2.70), 1.62 (d, J =6.84 Hz, 1.87), 1.55 (d, J =6.84 Hz, 3.08), 1.50 (d, J =6.59 Hz,1.46), 1.29 - 1.47 (m, 6.68), 0.87 (s, 9.20), 0.02 (s, 6.00).

Intermediate 14B (second eluting diastereomer)

Racemic 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (210 mg, 0.235 mmol, 38.2% yield) as a white solid.

LCMS ( m/z ): 895.6 [M + H] ⁺ ; 1.42 min retention time (Acquity UPLC CSH C18, 1.7 um, 50 mm x 2.1 mm column; 40°C; 3-95% gradient over 1.5 min, MeCN/10 mM ammonium bicarbonate in water adjusted to pH 10 with 25% ammonium hydroxide solution).

¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 7.80 (br. s., 2.29), 7.60 (br. s., 3.77), 7.43 (br. s., 3.64), 7.24 (br. s. , 3.24), 7.10 (s, 3.41), 6.81 (br.s., 3.41), 5.62 - 6.31 (br. s., 2.02), 5.35 - 5.61 (m, 3.60), 5.26 (br. s., 2.06 ), 4.57 - 4.84 (m, 11.14), 4.25 - 4.52 (m, 4.04), 4.11 (br. s., 1.46), 3.96 (t, J =5.96 Hz, 2.56), 3.84 (br. s., 0.87), 3.59 (br. s., 4.64), 2.85 (q, J =12.17 Hz, 0.97), 2.11 - 2.40 (m, 20.85), 2.06 (s, 5.95), 1.69 - 1.99 (m,8.53), 1.20 - 1.68 (m, 44.08), 0.96 (s, 10.71), 0.86 (s, 21.28), 0.14 (d, J =5.07 Hz, 7.03), 0.01 (d, J =3.80 Hz, 14.00).

Intermediate 15

4-Chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide

To 4-chloro-3-hydroxy-5-nitrobenzamide (942 mg, 4.35 mmol) dissolved in DMF (7 mL) was added Cs ₂ CO ₃ (1.559 g, 4.78 mmol) followed by 4-methoxybenzyl chloride (0.622 mL, 4.57 mmol). The reaction mixture was stirred at room temperature for 24 h. Under vigorous stirring, water (15 mL) was added dropwise and the resulting solid was stirred for 5 min, collected by filtration and rinsed with water to obtain the title compound (1.26 g, 3.74 mmol, 82% yield) as a light orange solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.80 (d, J=1.8 Hz, 1 H), 7.76 (d, J=1.8 Hz, 1 H), 7.43 (d, J=8.6 Hz, 2 H), 6.98 (d, J=8.6 Hz, 2 H), 6.13 (br. s., 1 H),5. 82 (br. s., 1 H), 5.25 (s, 2 H), 3.87 (s, 3 H); LCMS ( m/z ): 337.1 [M + H] ⁺ .

Intermediate 16

(E)-2,3-Dimethylbut-2-ene-1,4-diamine, 2-hydrochloride

Step 1: (E)-2,2'-(2,3-dimethylbut-2-ene-1,4-diyl)bis(isoindoline-1,3-dione)

To a solution of (E)-1,4-dibromo-2,3-dimethylbut-2-ene (29.5 g, 122 mmol) in DMF (244 mL) was added potassium phthalimide (45.2 g, 244 mmol). The white suspension was stirred at room temperature overnight. The reaction was poured into water (2 L) and the resulting white suspension was filtered. The filter cake was air dried (48 h) to give (E)-2,2'-(2,3-dimethylbut-2-ene-1,4-diyl)bis(isoindoline-1,3-dione) (37 g, 99 mmol, 81% yield) as a white solid. The solid was used without further purification. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 7.84 -7.93 (m, 4 H), 7.72 - 7.81 (m, 4 H), 4.39 (s, 4 H), 1.95 (s, 6 H).

Step 2: (E)-2,3-Dimethylbut-2-ene-1,4-diamine, 2-hydrochloride

To a mixture of (E)-2,2'-(2,3-dimethylbut-2-ene-1,4-diyl)bis(isoindoline-1,3-dione) (15.3 g, 40.9 mmol) in EtOH (332 mL) was added hydrazine monohydrate (6.01 mL, 123 mmol). The reaction was heated at 80°C. After 3 h, the reaction was cooled to room temperature. The thick white mixture was filtered, the filter cake was washed with ethanol, and the filtrate was concentrated to dryness. The resulting white solid from the filtrate was distributed between water (150 mL) and EtOAc (150 mL). The aqueous layer was concentrated to dryness to obtain a viscous yellow hued oil. The viscous oil was treated with 1 N HCl (250 mL) and EtOAc (250 mL). The white precipitate (by-product) that appeared was removed by filtration. The filtrate was transferred to a separatory funnel. The aqueous layer was separated; filtered to remove the remaining white solid and concentrated to dryness to obtain (E)-2,3-dimethylbut-2-ene-1,4-diamine, 2 hydrochloride (4.2 g, 22.45 mmol, 54.9% yield) as a greyish-pink solid. This material was used without further purification. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.23 (br. s., 6 H), 3.45 (q, J =5.75 Hz, 4 H), 1.83 (s, 6 H).

Intermediate 17

tert-Butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

Step 1: tert-Butyl (E)-(4-((2-amino-4-carbamoyl-6-methoxyphenyl)amino)but-2-en-1-yl)carbamate

To a 2-L round bottom flask was added tert-butyl (E)-(4-((4-carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)carbamate (25.8 g, 67.8 mmol) and methanol (484 mL). The orange heterogeneous solution was cooled to 0°C. After stirring at 0°C for 20 min, ammonium hydroxide solution (29% wt, 91 mL, 678 mmol) was added, followed by sodium sulfoxylate (85% wt, 70.0 g, 342 mmol) as a solution in water (194 mL). The flask was removed from the ice bath and stirred at room temperature. The heterogeneous mixture slowly changed color - from orange to off-white. After stirring at room temperature for 3 h, water (~800 mL) was added until a clear solution was obtained. The methanol was evaporated using reduced pressure. The white solid formed during evaporation was filtered off and washed twice with water (300 mL each). The solid was air dried for 16 h and then in a vacuum oven at 50 °C for 5 h. (E)-tert-butyl(4-((2-amino-4-carbamoyl-6-methoxyphenyl)amino)but-2-en-1-yl)carbamate (19.34 g, 54.1 mmol, 80% yield) was obtained as an off-white solid. The purity of this solid was 98% as judged by HPLC, LCMS and 1H NMR. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.62 (br. s., 1 H) 6.98 (br. s., 1H) 6.92 (t, J =5.45 Hz, 1 H) 6.87 (d, J =1.77 Hz, 1 H) 6.79 (d, J =1.77 Hz, 1 H)5.57 (qt, J =15.27, 5.23 Hz, 2 H) 4.67 (br. s., 2 H) 3.82 (br. s., 1 H) 3.76 (s, 3 H) 3.51 (dd, J =12.29, 5.70 Hz, 4 H) 1.37 (s, 9 H). LCMS ( m/z ): 351.1 (M+ H) ⁺ .

Step 2: tert-Butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

Into a 2-L round bottom flask was placed tert-butyl (E)-(4-((2-amino-4-carbamoyl-6-methoxyphenyl)amino)but-2-en-1-yl)carbamate (19.34 g, 55.2 mmol) and DMF (184 mL). This solution was cooled to 0°C. After stirring at 0°C for 20 min, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (44.2 mL, 44.2 mmol) was added as a ca. 1.0 M solution in dioxane. After stirring at 0°C for 10 min, the intermediate thiourea was completely formed. EDC (15.87 g, 83 mmol) and DIEA (28.9 mL, 166 mmol) were added. The reaction was allowed to warm to room temperature and stirred overnight (~14 h). To the heterogeneous reaction mixture was added a mixture of 250 mL saturated aqueous ammonium chloride and 750 mL water. This heterogeneous mixture was stirred at room temperature for 1 h. The solid was filtered off and rinsed twice with water (200 mL each time). The off-white solid was dried in a vacuum oven at 50 °C for 3 days. Tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (21.23 g, 41.5 mmol, 75% yield) was obtained as a white solid with a purity of ~100% as judged by LCMS, HPLC and ¹ H NMR. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.84 (br. s., 1 H) 8.00 (br. s., 1 H)7.67 (s, 1 H) 7.30 - 7.45 (m, 2 H) 6.86 - 7.00 (m, 1 H) 6.64 (s, 1 H) 5.54 -5 .80 (m, 2 H) 4.92 (d, J =4.82 Hz, 2 H) 4.61 (q, J =7.01 Hz, 2 H) 3.97 (s, 3 H)3.50 (br. s., 2 H) 2.18 (s, 3 H) 1.11 - 1.41 (m, 12 H). LCMS ( m/z ): 512.5 (M +H) ⁺ .

Intermediate 18

(E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

To a 1-L round bottom flask was added tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (21.23 g, 41.5 mmol), ethanol (234 mL) and tert-butyl methyl ether (96 mL). To this heterogeneous solution was added HCl (114 mL, 456 mmol) as a 4 M solution in dioxane. During the addition of HCl, the solution changed from heterogeneous to homogeneous with a clear yellow color. The reaction was stirred at room temperature overnight. By the next morning, a white solid had precipitated. More 4M HCl solution (15.56 mL, 62.2 mmol) was added and the mixture was stirred for an additional 9 h until the reaction was complete. The white solid was filtered off and rinsed with a 1:4 ethanol (200 mL)/TBME (800 mL) mixture. The resulting solid was dried in a vacuum oven overnight (50°C). (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (22.56 g, 44.2 mmol, 107% yield) was obtained as a white solid with a purity of 95% as judged by LCMS, HPLC and ¹ H NMR. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm12.88 (br. s., 1 H) 7.77 - 8.18 (m, 4 H) 7.68 (d, J =1.27 Hz, 1 H) 7.25 - 7.51 (m, 2 H) 6.68 (s, 1 H) 6.04 (dt, J =15. 52, 5.80 Hz, 1 H) 5.53 - 5.78 (m, 1 H)4.99 (d, J =5.32 Hz, 2 H) 4.61 (q, J =7.10 Hz, 2 H) 3.99 (s, 3 H) 3.27 - 3.57(m, 2 H) 2.19 (s, 3 H) 1.36 (t, J =7.10 Hz, 3 H). LCMS ( m/z ): 412.3 (M + H) ⁺ .

Intermediate 19

(E)-2-(4-amino-2,3-dimethylbut-2-en-1-yl)isoindoline-1,3-dione

Step 1: (3r,5r,7r)-1-((E)-4-Bromo-2,3-dimethylbut-2-en-1-yl)-1,3,5,7-tetraazaadamantan-1-ium bromide

To a solution of (E)-1,4-dibromo-2,3-dimethylbut-2-ene (13.59 g, 50.6 mmol) in DCM (200 mL) was added 1,3,5,7-tetraazaadamantane (7.09 g, 50.6 mmol) portionwise over 2 min. The reaction was stirred for 25 min and the resulting solid was filtered, rinsed with DCM and dried to obtain (3r,5r,7r)-1-((E)-4-bromo-2,3-dimethylbut-2-ene-1-yl)-1,3,5,7-tetraazaadamantane-1-azonia, bromide (16.7 g, 43.7 mmol, 86% yield) as a white solid. LCMS ( m/z ): 301.1 [M] ⁺ .

Step 2: (3r,5r,7r)-1-((E)-4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)-1,3,5,7-tetraazaadamantan-1-ium bromide

To a suspension of (3r,5r,7r)-1-((E)-4-bromo-2,3-dimethylbut-2-en-1-yl)-1,3,5,7-tetraazaadamantane-1-azonia, bromide (16.7 g, 43.7 mmol) in acetone (200 mL) was added Potassium 1,3-dioxoisoindolin-2-ide (8.09 g, 43.7 mmol). The reaction mixture was heated at 55 °C for 1.5 h. Over the next 2.5 h, the reaction was treated with additional potassium phthalimide until the starting material was consumed. The reaction mixture was removed from the heat source, stirred for 10 min, and then filtered while still warm. The solid was rinsed with acetone and dried to give 20.4 g of crude product. The crude product was stirred in cold water (ice bath) for 5 min. The solid was collected on a filter, rinsed with cold water, and dried to provide (3r,5r,7r)-1-((E)-4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)-1,3,5,7-tetraazaadamantane-1-azonia, bromide (10.3 g, 22.9 mmol, 52.6% yield) as a light yellow solid. LCMS ( m/z ): 368.2 [M] ⁺ .

Step 3: (E)-2-(4-amino-2,3-dimethylbut-2-en-1-yl)isoindoline-1,3-dione

To a suspension of (3r,5r,7r)-1-((E)-4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)-1,3,5,7-tetraazaadamantane-1-azonia, bromide (10.3 g, 22.97 mmol) in EtOH (100 mL) at room temperature was added concentrated hydrochloric acid (7.55 mL, 92 mmol). The color of the reaction mixture changed from light yellow to light orange. The reaction was heated at 80 °C for 55 min. The color changed to a darker orange over time. The reaction mixture was cooled to room temperature and saturated NaHCO ₃ solution was added to increase the pH of the solution (approximately 20 mL). The mixture was stirred for 5 min, diluted with 20 mL of water and extracted with 3:1 chloroform:EtOH (3x75 mL). The organic extract was dried over sodium sulfate, concentrated and dried to provide (E)-2-(4-amino-2,3-dimethylbut-2-en-1-yl)isoindoline-1,3-dione (5.77 g, 22.4 mmol, 98% yield) as a light brown solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.71 - 8.09 (m, 6 H), 4.25 (s, 2 H), 3.45 (s, 2 H), 1.95 (d, J =1.27 Hz, 3 H), 1.65 (d, J =1.27 Hz, 3 H). LCMS ( m/z ): 245.2 [M + H] ⁺ .

Intermediate 20

(E)-tert-Butyl (4-amino-2,3-dimethylbut-2-en-1-yl)carbamate

Step 1: tert-Butyl dichlorocarbamate

To a solution of tert-butyl carbamate (20.1 g, 172 mmol) in DCM (400 mL) was added calcium hypochlorite (technical grade, available chlorine 65%) (75 g, 343 mmol) at 0°C, followed by the addition of 6 M hydrochloric acid (143 mL, 858 mmol) dropwise over 35 min (internal temperature 5-10°C during addition). The resulting yellow suspension was then stirred for 20 min. The layers were separated, the organic layer was washed with water and brine, and dried over sodium sulfate. The solution was carefully concentrated under reduced pressure (23°C, 80 mbar) to provide tert-butyl dichlorocarbamate (33.4 g, 172 mmol, 100%) as a light yellow liquid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 1.56 (s, 9 H).

Step 2: tert-Butyl (E)-(4-chloro-2,3-dimethylbut-2-en-1-yl)carbamate

Nitrogen was bubbled through 300 mL of chloroform for 10 min. Then, 2,3-dimethylbut-1,3-diene (24.79 mL, 219 mmol) was added and the solution was cooled to 0 °C under a nitrogen atmosphere. A solution of tert-butyl dichlorocarbamate (41 g, 220 mmol) in chloroform (150 mL) was added over 80 min to produce a mixture of tert-butyl (E)-chloro(4-chloro-2,3-dimethylbut-2-en-1-yl)carbamate and tert-butyl (E)-(4-chloro-2,3-dimethylbut-2-en-1-yl)carbamate. After stirring in the ice bath for an additional 15 min, a freshly prepared aqueous sodium sulfite solution (3M, 219 mL, 657 mmol) was added dropwise rapidly at a rate to keep the internal temperature below room temperature (Caution: exothermic reaction with gas evolution). The ice bath was removed and the reaction was stirred for an additional 15 min. The layers were separated. The organic layer was washed with water and brine, dried over sodium sulfate, and concentrated to provide tert-butyl (E)-(4-chloro-2,3-dimethylbut-2-en-1-yl)carbamate (45.4 g, 184 mmol, 84% yield) as a white solid. ¹ H NMR (400 MHz, CHLOROFORM- d ) δ ppm 4.49 (br. s., 1 H), 4.10 (s, 2H), 3.80 (br. s., 2 H), 1.86 (s, 3 H), 1.82 (d, J =1.25 Hz, 3 H), 1.47 (s, 9H).

Step 3: tert-Butyl (E)-(4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)carbamate

To a solution of (E)-(4-chloro-2,3-dimethylbut-2-ene-1-yl)carbamic acid tert-butyl ester (40.4 g, 164 mmol) in DMF (300 mL) was added potassium phthalimide (30.4 g, 164 mmol) and the reaction mixture was stirred at room temperature for 3 h. The mixture was cooled in an ice/water bath and water (450 mL) was added to provide a thick precipitate. After stirring at room temperature for 10 min, the solid was filtered, rinsed with water, and dried to provide (E)-(4-(1,3-dioxoisoindoline-2-yl)-2,3-dimethylbut-2-ene-1-yl)carbamic acid tert-butyl ester (50.08 g, 137 mmol, 84% yield) as a white solid. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 7.87 (dd, J =5.40, 3.14 Hz, 2 H), 7.74 (dd, J =5.27, 3.01 Hz, 2 H), 4.41 - 4.48 (m, 1 H), 4.35 (s, 2 H), 3.78 (br. s., 2H), 1.97 (s, 3 H), 1.70 (d, J =1.25 Hz, 3 H), 1.47 (s, 9 H).

Step 4: tert-Butyl (E)-(4-amino-2,3-dimethylbut-2-en-1-yl)carbamate

Two identical reactions were set up in parallel. To a mixture of (E)-(4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-ene-1-yl)carbamic acid tert-butyl ester (25 g, 69.0 mmol) in ethanol (400 mL) was added hydrazine monohydrate (6.69 mL, 138 mmol). The mixture was stirred at 80 ° C for 4.5 h. After heating for 30 min, a thick precipitate began to form and stirring became difficult. The two reactions were combined and concentrated to remove ethanol and provide a white solid. This material was stirred in water (450 mL). 1M HCl (50 mL) and 6M HCl (14 mL) solutions were added to adjust the pH to ~5 and the suspension was stirred for 10 min. The solid was filtered off and rinsed with water. The aqueous filtrate was extracted with DCM (100 mL) to remove any impurities/color. The 1-hydroxy-2-(4-amino-2,3-dimethylbut-2-ene-1-yl) t-butyl carbamate (23.9g, ₁₁₁ mmol, 80% yield) of (E)-(4-amino-2,3-dimethylbut-2-ene-1-yl) t-butyl carbamate (23.9g, 111 mmol, 80% yield) was added to the 4-hydroxy-2-(4-amino-2,3-dimethylbut-2-ene-1-yl) carbamate (23.9g, 111 mmol, 80% yield) solution in 5 mL diethyl ether.Then the aqueous phase was adjusted to pH 13 with 1M sodium hydroxide and extracted with 3:1 CHCl 3:EtOH (3 x 300 mL).The organic layer merged through sodium sulfate drying, filtered, and concentrated to provide a light orange oil, which quickly solidifies.Grind this solid 5 min with 5% diethyl ether/heptane (200 mL), then filter and rinse with heptane (the 1st batch product).Concentrate the filtrate and stir in 5 mL diethyl ether.Rinse solid with minimum diethyl ether and filter to provide second batch product.Merge and dry in vacuum to provide (E)-(4-amino-2,3-dimethylbut-2-ene-1-yl) t-butyl carbamate (23.9g, 111 mmol, 80% yield) as off-white solid. ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 3.70 (s, 2 H), 3.24 (s, 2 H), 1.81 (d, J =1.00 Hz, 3 H), 1.73 (s, 3 H), 1.46 (s, 9 H). LCMS ( m/z ): 215.3 [M + H] ⁺ .

Intermediate 21

(E)-4-((4-Amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide, hydrochloride

Step 1: tert-Butyl (E)-(4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)carbamate

To a solution of (E)-(4-amino-2,3-dimethylbut-2-ene-1-yl)carbamic acid tert-butyl ester (1.92 g, 8.96 mmol) and 4-fluoro-3-nitrobenzamide (1.650 g, 8.96 mmol) in DMSO (25 mL) was added potassium carbonate (1.486 g, 10.75 mmol). The bright orange mixture was stirred at room temperature for 2 h. The mixture was added dropwise to rapidly stirred ice water (200 mL) and stirred for 1 h. The resulting precipitate was filtered, rinsed with water, and dried to provide (E)-(4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-ene-1-yl)carbamic acid tert-butyl ester (2.9 g, 7.5 mmol, 84% yield) as a bright yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.66 (d, J =2.28Hz, 1 H), 8.36 (t, J =5.32 Hz, 1 H), 7.99 (dd, J =8.87, 2.03 Hz, 2 H), 7.31 (br. s., 1 H), 7.02 (t, J =5 .70 Hz, 1 H), 6.92 (d, J =9.12 Hz, 1 H), 4.02 (d, J =5.07 Hz, 2 H), 3.60 (d, J =5.58 Hz, 2 H), 1.75 (s, 3 H), 1.68 (s, 3 H), 1.38 (s, 9 H).

Step 2: (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide, hydrochloride

To a suspension of tert-butyl (E)-(4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)carbamate (2.9 g, 7.66 mmol) in DCM (LARA updated) was added 4M HCl in dioxane (9.58 mL, 38.3 mmol). The reaction was stirred at room temperature for 2 h. The resulting solid was filtered, rinsed with DCM, and dried to afford (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide, hydrochloride (2.4 g, 7.28 mmol, 95% yield) as a bright yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm8.68 (d, J =2.28 Hz, 1 H), 8.44 (t, J =5.45 Hz, 1 H), 8.01 (dd, J =9.00, 2.15Hz, 5 H), 7.33 (br. s., 1 H), 6.90 (d, J =9. 12 Hz, 1 H), 4.10 (d, J =5.07 Hz, 2H), 3.48 (d, J =5.58 Hz, 2 H), 1.90 (s, 3 H), 1.72 (s, 3 H). LCMS ( m/z ): No significant [M + H] ⁺ .

Intermediate 22

(2S,3S)-2,3-diethoxybutane-1,4-diamine

Step 1: (4S,5S)-4,5-bis(azidomethyl)-2,2-dimethyl-1,3-dioxolane

To a solution of ((4S,5S)-2,2-dimethyl-1,3-dioxolane-4,5-diyl)bis(methylene)bis(4-methylbenzenesulfonate) (5.23 g, 11.11 mmol) in DMF (20 mL) was added sodium azide (2.89 g, 44.5 mmol). The mixture was stirred at 80 °C for 18 h. The mixture was diluted with water (100 mL) and extracted with EtOAc (2 x 100 mL). The organic phase was washed with water (2 x 100 mL), brine (100 mL), dried over magnesium sulfate and concentrated to provide (4S,5S)-4,5-bis(azidomethyl)-2,2-dimethyl-1,3-dioxolane (2.3 g, 10.8 mmol, 98% yield) as a clear oil. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 1.51 (s, 6 H), 3.30 - 3.43 (m, 2 H), 3.54 - 3.66 (m, 2 H), 4.10 (td, J =2.8, 1.3 Hz, 2 H). LCMS ( m/z ): No significant [M +H] ⁺ .

Step 2: (2S,3S)-1,4-diazidebutane-2,3-diol

To a solution of (4S,5S)-4,5-bis(azidomethyl)-2,2-dimethyl-1,3-dioxolane (2.3 g, 10.84 mmol) in THF (50 mL) was added p-toluenesulfonic acid (0.103 g, 0.542 mmol). The reaction mixture was heated at 60 °C for 18 h. The reaction mixture was cooled to room temperature and partitioned between EtOAc (50 mL) and water (30 mL). The aqueous phase was extracted with EtOAc (2 x 50 mL). The combined organic phases were washed with brine (30 mL), dried over magnesium sulfate, and concentrated. NMR analysis indicated that no reaction occurred. 1.25 M HCl in methanol (34.7 mL, 43.4 mmol) was added to the mixture. The reaction was heated at 60 °C for 18 h. The reaction mixture was concentrated to afford (2S,3S)-1,4-diazidobutane-2,3-diol (2.01 g, 10.5 mmol, 97% yield) as a light yellow clear oil. ¹ H NMR (400 MHz, CHLOROFORM- d ) δ ppm 3.35 - 3.59 (m, 4 H), 3.71 - 3.90 (m, 2 H).

Step 3: (2S,3S)-2,3-diethoxybutane-1,4-diamine

To a mixture of (2S,3S)-1,4-diazide butane-2,3-diol (2.01 g, 11.68 mmol) in DMF (50 mL) was added sodium hydride (1.167 g, 29.2 mmol) at 0 ° C. The mixture was stirred at room temperature for 5 min, and then iodoethane (2.36 mL, 29.2 mmol) was added. The mixture was stirred at room temperature for 18 h. The mixture was distributed between EtOAc (100 mL) and water (100 mL). The organic phase was washed with brine (3 x 30 mL), dried over magnesium sulfate and concentrated to provide a crude product (2S,3S)-1,4-diazide-2,3-diethoxybutane (2.47 g) as a clear oil. ^1H NMR (400 MHz, chloroform- d ) δ ppm 1.27 (t, J =7.0 Hz, 6 H), 3.31 - 3.47 (m, 4 H), 3.56 - 3.82 (m, 6H). A mixture of crude (2S, 3S)-1,4-diazide-2,3-diethoxybutane (2.47 g) and palladium on carbon (0.3 g, 2.8 mmol) in methanol (30 mL) was purged with nitrogen and exchanged with a hydrogen atmosphere (balloon). The mixture was stirred at room temperature for 18 h. Hydrogen was exchanged with nitrogen, the mixture was filtered through diatomaceous earth and concentrated to provide (2S, 3S)-2,3-diethoxybutane-1,4-diamine (1.86 g, 90% yield) as a clear oil. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 1.07 -1.13 (m, 6 H), 2.31 - 2.49 (m, 2 H), 2.57 - 2.68 (m, 2 H), 3.20 - 3.27 (m, 2H), 3.50 - 3.60 (m, 4 H).

Intermediate 23

4-Chloro-3-nitro-5-(trifluoromethyl)benzamide

To a solution of 4-chloro-3-nitro-5-(trifluoromethyl)benzoic acid (3.94 g, 14.62 mmol) in DCM (97 mL) was added oxalyl chloride (2.047 mL, 23.39 mmol) and 4 drops of DMF at room temperature. After stirring for 1 h, 30% ammonium hydroxide solution (9.49 mL, 73.1 mmol) was added and stirred for 18 h. The resulting white precipitate was filtered, first washed with water, then with DCM and dried to provide 4-chloro-3-nitro-5-(trifluoromethyl)benzamide (3.32 g, 12.36 mmol, 85% yield) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.77 (d, J =2.0 Hz, 1 H), 8.57 (d, J =1.8 Hz, 1 H), 7.97 (br. s., 2 H). LCMS ( m/z ): 269.1 [M + H] ⁺ .

Intermediate 24

Ethyl 3-(5-carbamoyl-2-fluoro-3-nitrophenyl)propionate

To 3-bromo-4-fluoro-5-nitrobenzamide (5 g, 18.25 mmol) in DMF (60.8 mL) was added tetra-n-butylammonium chloride (5.18 g, 18.25 mmol) and Pd(OAc) ₂ (0.418 g, 1.825 mmol). Nitrogen was bubbled through the mixture for 2 min, then 3,3-diethoxyprop-1-ene (8.69 mL, 54.7 mmol) and tributylamine (8.83 mL, 36.5 mmol) were added. The vessel was sealed and the mixture was heated at 125 °C for 16 h. The mixture was distributed between ethyl acetate (200 mL) and saturated aqueous ammonium chloride solution (200 mL). The aqueous phase was extracted again with ethyl acetate. The combined organic layers were washed with brine (2x150 mL), dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (120 g silica gel, 30-100% gradient of EtOAc/hexanes) to provide ethyl 3-(5-carbamoyl-2-fluoro-3-nitrophenyl)propanoate (1.77 g, 5.42 mmol, 29.7% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.48 (dd, J =6.7, 2.2 Hz, 1 H), 8.18 - 8.33 (m, 2 H), 7.71 (br. s., 1 H), 4.05 (q, J =7.3 Hz, 2 H), 2.94 - 3.06 (m, 2 H), 2.73 (t, J =7.5 Hz, 2 H), 1.15 (t, J =7.3 Hz, 3 H). LCMS ( m/z ): 285.1 [M + H] ⁺ .

Intermediate 25

2-Fluoro-1-((4-methoxybenzyl)oxy)-3-nitrobenzene

To the brown solution of 2-fluoro-3-nitrophenol (4.75 g, 30.2 mmol) in DMF (40 mL), cesium carbonate (10.84 g, 33.3 mmol) and 4-methoxybenzyl chloride (4.32 mL, 31.7 mmol) were added. The mixture was stirred at room temperature for 16 h. Water (150 mL) was added to the vigorously stirred reaction mixture and stirred for 10 min to produce a precipitate. The solid was filtered, rinsed with water, and dried to provide 2-fluoro-1-((4-methoxybenzyl)oxy)-3-nitrobenzene (8.1 g, 28.1 mmol, 93% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.63 - 7.75 (m, 2 H), 7.42 (d, J =8.62 Hz, 2 H), 7.36 (d, J =1.77 Hz, 1 H), 6.92 - 7.02 (m, 2 H), 5.21 (s, 2H), 3.77 (s, 3 H). LCMS ( m / z ): [M + H] ⁺ not observed.

Intermediate 26

(2R,3S)-2,3-Dimethoxybutane-1,4-diamine

Step 1: (2R,3S)-2,3-dimethoxysuccinic acid dimethyl ester

To a mixture of (2R,3S)-2,3-dihydroxysuccinic acid dimethyl ester (5.86 g, 32.9 mmol) and silver oxide (22.87 g, 99 mmol) was added iodomethane (41.1 mL, 658 mmol). The mixture was heated at 45 ° C for 6 h and kept at room temperature for 18 h. The mixture was filtered, washed with DCM and concentrated to provide the title compound as a clear oil (5.8 g, 28.3 mmol, 86% yield), which solidified after storage. ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 4.26 (s, 2 H) 3.76 (s, 6 H) 3.46 (s, 6 H).

Step 2: (2R,3S)-2,3-dimethoxybutane-1,4-diol

A solution of (2R,3S)-2,3-dimethoxysuccinic acid dimethyl ester (5.1 g, 24.73 mmol) in THF (30 mL) was added to a mixture of LAH (2.065 g, 54.4 mmol) in THF (150 mL) at 0 ° C. The mixture was warmed to room temperature for 2 h. The reaction was quenched with saturated sodium sulfate solution (9.1 mL). The mixture was filtered, dried over magnesium sulfate and concentrated to provide the title compound (3.6 g, 24.0 mmol, 97% yield) as a clear oil. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 4.51 (t, J =5.58 Hz, 2 H) 3.51 - 3.59 (m, 2 H) 3.38 - 3.45 (m, 2 H) 3.32 (s, 6 H) 3.19 - 3.26 (m, 2 H).

Step 3: (2R,3S)-2,3-dimethoxybutane-1,4-diyl bis(4-methylbenzenesulfonate)

To a solution of (2R,3S)-2,3-dimethoxybutane-1,4-diol (3.3 g, 21.97 mmol) in pyridine (40 mL) was added TsCl (12.57 g, 65.9 mmol) at -78 °C. The mixture was allowed to warm to room temperature and stirred for 18 h. Water (150 mL) was added and the mixture was cooled to 0 °C for 2 h. The resulting precipitate was filtered, rinsed with water and dried to provide the title compound (7.52 g, 16.4 mmol, 74.6% yield) as a white solid. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 7.69 - 7.86 (m, 4 H) 7.35 - 7.44 (m, 4 H) 4.08 - 4.33 (m, 4 H) 3.33- 3.45 (m, 2 H) 3.25 (s, 6 H) 2.49 (s, 6 H).

Step 4: (2R,3S)-1,4-diazide-2,3-dimethoxybutane

To a solution of bis(4-methylbenzenesulfonic acid) (2R,3S)-2,3-dimethoxybutane-1,4-diyl ester (7.52 g, 16.40 mmol) in DMF (40 mL) was added sodium azide (4.26 g, 65.6 mmol). The mixture was stirred at 80 °C for 18 h. The mixture was diluted with water (200 mL) and extracted with EtOAc (2 x 200 mL). The organic phase was washed with water (2 x 200 mL) and brine (100 mL), dried over magnesium sulfate and concentrated to provide the title compound (3.16 g, 15.8 mmol, 96% yield) as a clear oil. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 3.60 - 3.68 (m, 2 H) 3.48 (s, 6 H) 3.37 - 3.45 (m, 4 H).

Step 5: (2R,3S)-2,3-dimethoxybutane-1,4-diamine

To a mixture of (2R, 3S)-1,4-diazide-2,3-dimethoxybutane (3.16 g, 15.8 mmol) and palladium on carbon (0.672 g, 6.31 mmol) in methanol (30 mL) was added hydrogen (balloon). The mixture was stirred at room temperature for 60 h. After removing the hydrogen, the mixture was filtered through diatomaceous earth and concentrated to provide the title compound (2.33 g, 15.7 mmol, 100% yield) as a clear oil. ¹ H NMR (400 MHz, chloroform- d ) δ ppm 3.49 (s, 6 H) 3.25 - 3.35 (m, 2 H) 2.83 - 2.96 (m, 4 H).

Intermediate 27

((1S,2S)-Cyclopropane-1,2-diyl)dimethylamine, 2-hydrochloride

Step 1: (1S,2S)-cyclopropane-1,2-dicarboxamide

To a 250-mL round-bottom flask was added (1S,2S)-cyclopropane-1,2-dicarboxylic acid diethyl ester (38 g, 204 mmol) and ammonium hydroxide solution (28% wt aqueous solution; 380 mL, 3035 mmol). The mixture was stirred at 25 °C for 48 h. The mixture was filtered and the filter cake was subsequently washed with EtOAc (100 mL). The solid was dried under vacuum to obtain (1S,2S)-cyclopropane-1,2-dicarboxamide (14.5 g, 108 mmol, 53% yield) as a white solid. ¹ H NMR (300 MHz, DMSO- d ₆ ) δ ppm 7.65 (s, 2 H), 6.90 (s, 2 H), 1.86 (m, 2 H), 0.97 (m, 2 H).

Step 2: Di-tert-butyl (((1S,2S)-cyclopropane-1,2-diyl)bis(methylene))dicarbamate

To a mixture of (1S,2S)-cyclopropane-1,2-dicarboxamide (14.5 g, 113 mmol) and THF (300 mL) was added LiAlH ₄ (17.18 g, 453 mmol) in portions at 0°C. Then, the mixture was stirred at 25°C for 48 h. The mixture was quenched by adding crushed ice (200 g) at 0°C. The mixture was filtered and the filtrate was used directly in the next step. LiOH (10.52 g, 4.39 mmol) and water (200 mL) were added to the filtrate. Boc anhydride (56.1 mL, 242 mmol) was added and the mixture was stirred at room temperature overnight. Then, the reaction mixture was extracted three times with DCM (100 mL x 3). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by normal phase silica gel chromatography (80 g silica gel, 1:4 EtOAc/petroleum ether) to provide di-tert-butyl (((1S,2S)-cyclopropane-1,2-diyl)bis(methylene))dicarbamate (10 g, 31.6 mmol, 29% yield over two steps) as a colorless oil. ¹ H NMR (300 MHz, methanol- d ₄ ,) δ ppm 3.03 (m, 2 H), 2.85 (m, 2 H), 0.83 (m, 2 H), 0.41 (m, 2 H).

Step 3: ((1S,2S)-cyclopropane-1,2-diyl)dimethylamine, 2-hydrochloride

To a 500-mL round-bottom flask was added di-tert-butyl (((1S,2S)-cyclopropane-1,2-diyl)bis(methylene))dicarbamate (10 g, 33.3 mmol) and HCl (4 M in 1,4-dioxane, 100 mL, 400 mmol). After stirring at 25 °C for 30 min, the mixture was concentrated under reduced pressure. The residue was then dissolved in water (100 mL) and lyophilized. ((1S,2S)-cyclopropane-1,2-diyl)dimethylamine, 2-hydrochloride (5.3 g, 29.1 mmol, 87% yield) was obtained as an off-white solid. ¹ H NMR (400 MHz, methanol- d ₄ ,) δ ppm 3.05 (m, 2 H), 2.81 (m, 2 H), 1.27 - 1.18 (m, 2 H), 0.87 - 0.79 (m, 2 H). LCMS ( m / z ): 101.2 [M + H] ⁺ , UV peak not observed.

Example 1

(E)-1-((E)-4-((Z)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

Step 1: (E)-1-(4-((4-Carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide hydrochloride (535 mg, 1.280 mmol) in EtOH (5 mL) was added triethylamine (471 mg, 4.65 mmol) and 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (400 mg, 1.164 mmol). The reaction vessel was sealed and heated at 120° C. for 20 h. After cooling, an orange solid precipitated from the dark solution. The solid was washed with EtOAc and dried to provide (E)-1-(4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (457 mg, 0.664 mmol, 57.0% yield). The reaction was repeated three times to provide 1.37 g of the title compound. LCMS m/z = 689 [M+H] ⁺ .

Step 2: (E)-1-(4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

(E)-1-(4-((4-Carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (1.05 g, 1.525 mmol) was suspended in MeOH (16 mL) and 28% ammonium hydroxide (5.17 mL, 38.1 mmol). After 5 min stirring, a solution of sodium sulfoxylate (1.593 g, 9.15 mmol) in water (4.00 mL) was added and then stirred at room temperature for 2 h.

EtOAc was added and the organic layer was washed with water and brine. Then, the organic phase was dried and concentrated to obtain a crude product (E) -1- (4- ((2- amino -4- formamyl -6- (3- morpholino propoxy) phenyl) amino) but-2-ene-1-yl) -2- (1- ethyl -3- methyl -1H- pyrazole -5- formamido) -1H- benzo [d] imidazole -5- carboxamide (330 mg, 0.501 mmol, 32.9% yield) as an off-white solid. The crude material was used without further purification. LCMS m/z = 659 [M + H] ⁺ .

Step 3: (E)-2-amino-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide

To a solution of (E)-1-(4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (330 mg, 0.501 mmol) in MeOH (15 mL) was added cyanogen bromide (159 mg, 1.503 mmol) and the reaction mixture was stirred at room temperature for 3 h. Precipitation of the product was achieved by addition of EtOAc and subsequent stirring for 1 h. The solid was filtered, washed with EtOAc and dried to afford (E)-2-amino-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide (284 mg, 0.416 mmol, 83% yield) as a light brown solid. The material was used without further purification. LCMS m/z = 684 [M + H] ⁺ .

Step 4: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-2-amino-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide (260 mg, 0.380 mmol) in DMF (4 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (117 mg, 0.760 mmol) and 1H-benzo[d][1,2,3]triazol-1-ol hydrate (58.2 mg, 0.760 mmol) at room temperature. 3-[(2 ... The crude product was further purified by silica gel chromatography (ISCO 24 g column, 0-35% gradient of MeOH in DCM) to afford after removal of solvent (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide (140 mg, 0.171 mmol). ¹ H NMR (Methanol - d ₄ , 600 MHz): δ ppm 7.96 (s, 1 H), 7.71 (dd, J =8.3, 1.6 Hz, 1 H), 7.56 (s, 1 H), 7.35 (d, J =8.4 Hz, 1 H), 7.27 (s, 1 H), 6.62 (s, 1 H ), 6.55 (s, 1 H), 5.95 (dt, J =15.5, 5.1 Hz, 1 H), 5.76-5.83 (m, 1 H), 5.06 (br d, J =4.6 Hz, 2 H), 4.86 (br d, J =5.3 Hz, 2 H), 4.63(s, 2 H), 4.5 6 (q, J =7.0 Hz, 2 H), 3.99 (t, J =6.1 Hz, 2 H), 3.64 (br t, J =4.2Hz, 4 H), 2.43-2.48 (m, 2 H), 2.40 (br s, 4 H), 2.21 (s, 3 H), 2.19 (s, 3 H),1.75-1.81 (m, 2 H) , 1.37 (t, J =7.1 Hz, 3 H), 1.32 (t, J =7.2 Hz, 3 H). LCMS m/z = 820.9 [M + H] ⁺ .

Step 5: (E)-1-((E)-4-((Z)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide (52 mg, 0.063 mmol) in DMF (2 mL) was added cesium carbonate (62.0 mg, 0.190 mmol) and iodomethane (9.91 µl, 0.159 mmol). The reaction mixture was stirred at room temperature for 12 h. The solvent was evaporated and the residue was purified by silica gel chromatography (0-25% gradient of MeOH/DCM, silica gel column 12 g) to obtain the clean product (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (44 mg, 0.052 mmol, 82% yield). ¹ H NMR (DMSO- d ₆ , 600 MHz): δ ppm 8.07 (br s, 1 H), 7.80 (br d, J =8.3 Hz, 1 H), 7.74 (br s, 1 H), 7.48 (br d, J =8.4Hz, 1 H), 7.41 (s, 1 H), 6.29-6. 44 (m, 2 H), 5.83-5.99 (m, 1 H), 5.60-5.76 (m, 1 H), 4.81-4.94 (m, 2 H), 4.75 (br d, J =5.1 Hz, 2 H), 4.38-4.55 (m, 4 H), 4.06 (br s, 2 H), 3.54 (br s, 3 H), 3.45-3.59 (m, 7 H), 2.25-2.30 (m, 2 H), 2.15-2.37 (m, 4 H), 2.11 (br d, J =7.0 Hz, 6 H), 1.72 (br s, 2 H), 1.19-1.24 (m, 3 H), 1.14-1.26 (m, 3 H). LCMS m/z = 848 [M + H] ⁺ .

The compounds prepared by the above method may exist in the form of tautomers/isomers, such as (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

Example 2

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)butyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: tert-Butyl (4-((4-carbamoyl-2-nitrophenyl)amino)butyl)carbamate

A mixture of tert-butyl (4-aminobutyl)carbamate (5.00 g, 26.6 mmol), 4-fluoro-3-nitrobenzamide (4.89 g, 26.6 mmol) and _K2CO3 (4.04 g, 29.2 mmol) in DMSO (25 mL) was stirred at 70°C for 2 _h . The reaction was cooled to room temperature and slowly diluted with 125 mL of water via an addition funnel. The resulting solid was isolated by filtration, dried and placed in a vacuum oven at 56°C for 3 days to provide the title compound (9.2 g, 26.1 mmol, 98% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.67 (d, J =2.02 Hz, 1 H) 8.40 (t, J =5.43 Hz, 1 H) 8.01 (d, J =6.82 Hz, 2 H) 7.30 (br. s., 1 H) 7.12 (d, J =9.09 Hz, 1 H) 6.87 (br. s., 1 H) 3.42 (q, J =6.57 Hz, 2 H) 2.91 - 3.01 (m, 2 H) 1.60 (d, J =6.57 Hz, 2 H) 1.43 - 1.54 (m, 2 H) 1.38 (s, 9 H). LCMS [M+H] ⁺ = 353.

Step 2: tert-Butyl (4-((2-amino-4-carbamoylphenyl)amino)butyl)carbamate

A 500 mL round bottom flask was charged with tert-butyl (4-((4-carbamoyl-2-nitrophenyl)amino)butyl)carbamate (9.2 g, 26.1 mmol), 10% Pd/C (0.920 g, 8.64 mmol) (Degussa wet type), EtOH (100 mL) and MeOH (100 mL). The flask was evacuated and placed under a hydrogen balloon with stirring. A condenser was placed above the flask and a hydrogen balloon was placed on the condenser. The mixture was stirred at room temperature for 20 h, then the flask was evacuated and the suspension was filtered through a diatomaceous earth bed using EtOH as a washing aid. The filtrate was concentrated under vacuum and placed under high vacuum to provide the title compound (8.4 g, 26.1 mmol, 100% yield) as a black solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm7.44 (br. s., 1 H) 7.04 - 7.15 (m, 2 H) 6.85 (t, J =5.43 Hz, 1 H) 6.74 (br.s., 1 H) 6.37 (d, J =8.08 Hz, 1 H) 4.89 (t, J =5.18 Hz, 1 H) 4.60 (br. s., 2 H)3.07 (q, J =6.48 Hz, 2 H) 2.97 (q, J =6.40 Hz, 2 H) 1.45 - 1.64 (m, 4 H) 1.39 (s, 9 H). LCMS [M + H] ⁺ = 323.1.

Step 3: tert-Butyl (4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)butyl)carbamate, hydrobromide

Tert-butyl (4-((2-amino-4-carbamoylphenyl)amino)butyl)carbamate (8.40 g, 26.1 mmol) was dissolved in MeOH (110 mL) and a 5 M solution of cyanogen bromide in _CH3CN (5.73 mL, 28.7 mmol) was added via syringe. The dark reaction was capped and stirred at room temperature for 15 h. The reaction was concentrated in vacuo and placed under high vacuum to provide the title compound as a dark solid (11.17 g, 26.1 mmol, 100% yield). ¹ HNMR (400 MHz, DMSO- d ₆ ) δ ppm 12.85 (br. s., 1 H) 8.74 (br. s., 2 H) 8.08 (br.s., 1 H) 7.80 - 7.90 (m, 2 H) 7.64 (d, J =8.34 Hz, 1 H) 7.44 (br. s., 1 H)6.89 (t, J =5.56 Hz, 1 H) 4.15 (t, J =7.20 Hz, 2 H) 2.96 (q, J =6.32 Hz, 2 H)1.66 (d, J =7.07 Hz, 2 H) 1.42 - 1.50 (m, 2 H) 1.38 (s, 9 H). LCMS [M+H] ⁺ = 348.1.

Step 4: tert-Butyl (4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)carbamate

A mixture of tert-butyl (4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)butyl)carbamate, hydrobromide (11.17 g, 26.1 mmol), 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (4.82 g, 31.3 mmol), HATU (11.90 g, 31.3 mmol), DIPEA (18.22 mL, 104 mmol), and HOBt (1.997 g, 13.04 mmol) in DMF (100 mL) was stirred at room temperature for 21 h. The reaction was diluted with 300 mL of water and 300 mL of EtOAc, transferred to a separatory funnel, the layers separated, and the aqueous layer extracted with EtOAc (2 x 150 mL). The combined EtOAc layers were washed with saturated NH ₄ Cl (2x 200 mL), water (1 x 200 mL), and brine (2 x 200 mL). The organic layer was dried over Na ₂ SO ₄ , filtered, concentrated in vacuo, and placed under high vacuum. The solid was purified via chromatography on silica gel (ISCO ^® Combiflash, 0-20% MeOH: DCM, 330 g column, loaded in 50 mL DCM). The desired fractions were combined, concentrated in vacuo, and placed under high vacuum to provide the title compound (9.53 g, 19.71 mmol, 76% yield) as a purple solid. ¹ HNMR (400 MHz, DMSO- d ₆ ) δ ppm 12.85 (s, 1 H) 8.01 (br. s., 2 H) 7.81 (d, J =8.34 Hz, 1 H) 7.59 (d, J =8.34 Hz, 1 H) 7.36 (br. s., 1 H) 6.80 - 6.86 ( m, 1H) 6.68 (s, 1 H) 4.64 (q, J =6.82 Hz, 2 H) 4.23 (t, J =6.44 Hz, 2 H) 2.98 (d, J =5.81 Hz, 2 H) 2.19 (s, 3 H) 1.76 (d, J =6.57 Hz, 2 H) 1.40 - 1.48 (m, 2 H)1.30 - 1.40 (m, 13 H). LCMS [M + H] ⁺ = 484.3.

Step 5: 1-(4-aminobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

An ice-cooled 500 mL round bottom flask containing tert-butyl (4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)carbamate (9.53 g, 19.71 mmol) was treated with 4 M HCl in 1,4-dioxane (42.0 mL, 168 mmol). The ice bath was removed and the purple slurry was stirred at room temperature for 2.5 h. The reaction was then concentrated in vacuo, placed under high vacuum, and the resulting solid was placed in a vacuum oven at 50 °C for 15 h and cooled under high vacuum to provide the impure title compound (11.89 g, assumed 19.7 mmol, 100% yield) as a gray solid also containing 1,4-dioxane. The material was used as is without further purification. ¹ H NMR (400MHz, DMSO- d ₆ ) δ ppm 12.91 (br. s, 1 H) 8.03 (d, J =1.26 Hz, 2 H) 7.77 - 7.87(m, 4 H) 7.62 (d, J =8.34 Hz, 1 H) 7.38 (br. s., 1 H) 6.70 (s, 1 H) 6-5 (br.s, 1 H), 4.63 (q, J =7.07 Hz, 2 H) 4.28 (t, J =6.57 Hz, 2 H) 2.77 - 2.87 (m, 2H) 2.20 (s, 3 H) 1.81 - 1.91 (m, 2 H) 1.52 - 1.60 (m, 2 H) 1.38 (t, J =7.07Hz, 3 H). LCMS [M + H] ⁺ = 384.2.

Step 6: 4-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)amino)-3-methoxy-5-nitrobenzoic acid methyl ester

A 250 mL 3-neck round bottom flask equipped with a condenser, large stirring bar and internal thermometer was charged with 1-(4-aminobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (9.38 g, 20.55 mmol) and methyl 4-chloro-3-methoxy-5-nitrobenzoate (5.048 g, 20.55 mmol). DMSO (50 mL) was added followed by DIPEA (17.95 mL, 103 mmol) and the dark suspension was heated at 100 °C for approximately 24 h, cooled, and added dropwise to 500 mL of stirring water. After the addition was complete, the resulting orange suspension was stirred for 20 min and filtered. The separated orange-red paste was washed with water and hexanes, dried in a Buchner funnel, and then dried in a vacuum oven at 56 °C for 20 h. The reddish solid was then triturated with _Et20 (60 mL) and isolated by filtration. The trituration and filtration were repeated. The resulting solid was placed in a vacuum oven at 56°C for 3 days to provide the title compound (11.17 g, 18.85 mmol, 92% yield) as a reddish solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.78 (br. s., 1 H) 8.12 (s, 1H) 7.99 (s, 1 H) 7.93 (d, J =7.53 Hz, 2 H) 7.79 (d, J =8.28 Hz, 1 H) 7.53 (d, J =7.78 Hz , 1 H) 7.36 (s, 1 H) 7.31 (br. s., 1 H) 6.60 (s, 1 H) 4.60 (d, J =7.03Hz, 2 H) 4.23 (br. s., 2 H) 3.84 (s, 3 H) 3.80 (s, 3 H) 3.53 (d, J =5.77 Hz, 2H) 2.15 (s, 3 H) 1.82 (br. s., 2 H) 1.62 (br. s., 2 H) 1.35 (t, J =7.03 Hz, 3H). LCMS [M + H] ⁺ = 711.6.

Step 7: 3-amino-4-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)amino)-5-methoxybenzoic acid methyl ester

Methyl 4-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)amino)-3-methoxy-5-nitrobenzoate (5.0 g, 8.44 mmol) was mostly dissolved in DMF (50 mL) in a 250 mL round bottom flask at room temperature with stirring. Raney nickel (Raney 2800 nickel in water, approximately 10 mL slurry, Aldrich) was added and a condenser was placed on top of the flask. A 3-way stopcock adapter with a hydrogen balloon was placed over the condenser and the apparatus was evacuated, filled with hydrogen, evacuated, and finally filled with hydrogen. The reaction was heated at 70 °C for 7 h. An additional 8 mL of Raney nickel was added and the reaction was heated at 70 °C for 14 h. The reaction was cooled and filtered through celite while washing with DMF. The filtrate containing the desired product, a solution of approximately 100 mL DMF and 20 mL water from the Raney nickel slurry, was used directly as a solution in the next reaction. Assumed quantitative yield. LCMS [M + H] ⁺ = 563.4.

Step 8: 2-amino-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid methyl ester, hydrobromide

Methyl 3-amino-4-((4-(5-carbamoyl-2-(1-ethyl-3-methyl- _1H -pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)amino)-5-methoxybenzoate (solution in DMF/water from the previous step) was treated with 5 M cyanogen bromide in CH 3 CN (1.875 mL, 9.37 mmol) and the resulting solution was stirred at room temperature for 22 h. The reaction was concentrated in vacuo and placed under high vacuum to afford a brown semisolid. The semisolid was triturated with EtOAc, stirred vigorously for 30 min, and the resulting solid was isolated by filtration and dried in a Buchner funnel to afford the impure title product as a brown solid (5.08 g). This impure material was used without purification. LCMS [M + H] ⁺ = 588.5.

Step 9: 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-5-carboxylic acid methyl ester

A mixture of methyl 2-amino-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate, hydrobromide (5.073 g, 7.59 mmol), 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (1.287 g, 8.35 mmol), HATU (3.46 g, 9.11 mmol) and DIPEA (3.98 mL, 22.76 mmol) in DMF (30 mL) was stirred at room temperature for 17 h. The reaction was concentrated in vacuo and the resulting residue was triturated with water (100 mL) and stirred for 30 min. The resulting suspension was filtered and partially dried in a Buchner funnel to provide a dark brown solid. Solid is dissolved in 150 mL of 10% IPA: chloroform mostly, diluted with water and filtered.Then, separate filtrate layer, and through Na ₂ SO ₄ dry organic layer, filter, concentrate and be placed under high vacuum to provide brown solid.Adopt warm 10% IPA: chloroform (100 mL) grinding solid and filter.Separate filtrate layer, through Na ₂ SO ₄ dry organic layer, filter, add in initial brown solid, concentrate and be placed under high vacuum in vacuum.Via chromatography on silica gel (Biotage ^® Isolera, 120 gm Gold post, through 30 min 0-10% MeOH: DCM, as the solution in DCM/MeOH load) upper purification of this solid.Merge required product fraction, concentrate, and be placed under high vacuum to provide light brown solid. The solid was triturated with DCM (50 mL) and isolated by filtration and placed in a vacuum oven at 56 °C for 30 h to afford methyl 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate (1.0 g, 1.4 mmol, 18% yield) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm12.89 (s, 1 H) 12.82 (s, 1 H) 7.90 - 8.01 (m, 2 H) 7.70 - 7.81 (m, 2 H) 7.53 (d, J =8.28 Hz, 1 H) 7.30 - 7.40 (m, 2 H ) 6.59 (d, J =5.02 Hz, 2 H) 4.50 - 4.64(m, 4 H) 4.38 (br. s., 2 H) 4.27 (br. s., 2 H) 3.87 (d, J =3.76 Hz, 6 H) 2.10(s, 6 H) 1.86 (br. s., 4 H) 1.23 - 1.39 (m, 6H). LCMS [M + H] ⁺ = 724.5.

Step 10: 1-(4-(5-Carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

To a suspension of methyl 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate (550 mg, 0.760 mmol) in MeOH (11 mL) and water (11 mL) was added NaOH (304 mg, 7.60 mmol). The reaction was stirred at room temperature overnight. The MeOH was removed in vacuo and the resulting solution was treated with 1 N HCl until pH ~3. The resulting slurry was filtered and the filter cake was dried in a vacuum oven to provide 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid (650 mg, 0.687 mmol, 90% yield) as a white solid. The compound was used in the next step without further purification. LCMS m/z = 710 [M +H] ⁺ .

Step 11: 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

To a solution of 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid (320 mg, 0.338 mmol) and HATU (154 mg, 0.406 mmol) in DMF (3.381 mL) was added DIEA (295 µL, 1.691 mmol). After 20 min, ammonium chloride (54.3 mg, 1.014 mmol) was added to the reaction mixture and the reaction was stirred at room temperature for 3 days. Additional HATU (50 mg, 0.132 mmol) and DIEA (58.9 µl, 0.338 mmol) were added. The reaction was stirred at room temperature for 10 min and ammonium chloride (18.26 mg, 0.338 mmol) was added. For complete reaction, HOBt hydrate (51.7 mg, 0.338 mmol) was added and the reaction was stirred at room temperature for 90 min. The reaction was dry loaded onto silica gel and purified by silica gel chromatography (ISCO-Rf 12g post, 0%-30% gradient of MeOH/DCM) to provide 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-formamido)-1H-benzo [d] imidazoles-1-yl) butyl)-2-(1-ethyl-3-methyl-1H-pyrazoles-5-formamido)-7-methoxyl-1H-benzo [d] imidazoles-5-carboxamide (175 mg, 0.244 mmol, 72.3% yield) as white solid. LCMS m/z = 709 [M+ H] ⁺ .

Step 12: (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)butyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (100 mg, 0.141 mmol) in DMF (4 mL) was added cesium carbonate (138 mg, 0.423 mmol) and iodomethane (50.1 mg, 0.353 mmol, 100 uL stock (220 uL iodomethane in 780 uL DMF). The reaction was stirred at room temperature for 2 h, dry loaded onto silica gel and purified by silica gel chromatography (ISCO-Rf, 12 g column, 0%-30% gradient of MeOH/DCM) to afford 100 mg solid (by LCMS, ~93% pure). The resulting residue (100 mg) was dissolved in MeOH, dry loaded onto silica gel and purified again (ISCO-Rf, 12 g column, 0%-20% gradient of MeOH/DCM). Pure fractions were combined and concentrated to dryness to provide (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo [d] imidazole-1-yl)butyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo [d] imidazole-5-carboxamide (33 mg, 0.044 mmol, 31.4% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.98 - 8.13 (m, 3 H)7.84 (dd, J =8.36, 1.52 Hz, 1 H) 7.69 (d, J =1.01 Hz, 1 H) 7.58 (d, J =8.62 Hz, 1 H) 7.47 (d, J =14.19 Hz, 2 H) 7.4 2 (s, 1 H) 6.46 (d, J =6.84 Hz, 2 H) 4.42 -4.55 (m, 4 H) 4.11 - 4.28 (m, 4 H) 3.83 (s, 3 H) 3.51 (s, 3 H) 3.47 (s, 3 H)2.12 (s, 6 H) 1.74 (br. s., 4 H) 1.24 (td, J =7.10, 1.52 Hz, 6 H). LCMS m/z =737 [M + H] ⁺ .

Example 3

(5aE,21E)-8-ethyl-5,10,18,22-tetramethyl-7,20-dioxo-5,7,8,11,12,13,14,15,20,22,28,29,30,31-tetradecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-l][1,3,6,15,17]pentaazacyclohenicosine-3,24-dicarboxamide

Step 1: 3-Methyl-1-(5-(trimethylsilyl)pent-4-yn-1-yl) -1H -pyrazole-5-carboxylic acid ethyl ester

A mixture of ethyl 3-methyl- 1H -pyrazole-5-carboxylate (22 g, 143 mmol), (5-chloropent-1-yn-1-yl)trimethylsilane (24.94 g, 143 mmol), K ₂ CO ₃ (39.4 g, 285 mmol) and DMF (4 mL) was stirred at 60° C. under a nitrogen atmosphere overnight. The mixture was then dissolved in DCM and washed with water. The organic phase was dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure, and purified by column chromatography on silica gel (petroleum ether/EtOAc=10:1) to provide ethyl 3-methyl-1-(5-(trimethylsilyl)pent-4-yn-1-yl) -1H -pyrazole-5-carboxylate (12.5 g, 42.7 mmol, 30% yield) as a colorless oil. LCMS [M + H] ⁺ = 293.

Step 2: 3-Methyl-1-(pent-4-yn-1-yl) -1H -pyrazole-5-carboxylic acid ethyl ester

A mixture of ethyl 3-methyl-1-(5-(trimethylsilyl)pent-4-yn-1-yl) -1H -pyrazole-5-carboxylate (37.7 g, 129 mmol), K ₂ CO ₃ (44.5 g, 322 mmol) and EtOH (800 mL) was stirred at room temperature overnight. Then, the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM, washed with water, dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to provide ethyl 3-methyl-1-(pent-4-yn-1-yl) -1H -pyrazole-5-carboxylate (20 g, 91 mmol, 70.4% yield) as a colorless oil. LCMS [M + H] ⁺ = 221.

Step 3: 1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid benzyl ester

A mixture of 1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid (20 g, 130 mmol), (bromomethyl)benzene (22.2 g, 130 mmol), _K2CO3 (26.9 g, 195 mmol) and DMF (200 _mL ) was stirred at 60°C overnight. The mixture was then dissolved in DCM, washed with water, dried over anhydrous _{Na2SO4 ,} filtered, concentrated under reduced pressure, and purified by column chromatography on silica gel (petroleum ether/EtOAc = 10:1) to provide 1-ethyl-3-methyl-pyrazole-5-carboxylic acid benzyl ester (31.4 g, 129 mmol, 99% yield) as a colorless oil. LCMS [M+H] ⁺ = 245.

Step 4: 1-ethyl-4-iodo-3-methyl- 1H -pyrazole-5-carboxylic acid benzyl ester

A mixture of 1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid benzyl ester (31.6 g, 129 mmol), 1-iodopyrrolidine-2,5-dione (34.9 g, 155 mmol) and DMF (400 mL) was stirred at 90 ° C for 2 days. Then, the mixture was cooled to room temperature, dissolved in DCM, and washed with saturated sodium _thiosulfate aqueous solution. The organic layer was dried over anhydrous _Na2SO4 , filtered, concentrated under reduced pressure, and purified by column chromatography (petroleum ether/EtOAc = 10: 1) to provide 1-ethyl-4-iodo-3-methyl- 1H -pyrazole-5-carboxylic acid benzyl ester (42.6 g, 115 mmol, 89% yield). LCMS [M + H] ⁺ = 371.

Step 5: Benzyl 4-(5-(5-(ethoxycarbonyl)-3-methyl- 1H -pyrazol-1-yl)pent-1-yn-1-yl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylate

A mixture of ethyl 3-methyl-1-(pent-4-yn-1-yl) -1H -pyrazole-5-carboxylate (10.0 g, 45.4 mmol), benzyl 1-ethyl-4-iodo-3-methyl- 1H -pyrazole-5-carboxylate (16.8 g, 45.4 mmol), copper (I) iodide (0.864 g, 4.54 mmol), bis (triphenylphosphine) palladium (II) chloride (0.319 g, 0.454 mmol) and _Et3N (200 mL) was stirred under nitrogen atmosphere overnight at 60° C. The mixture was then dissolved in DCM and washed with water. _The organic phase was dried over anhydrous _Na2SO4 , filtered, concentrated under reduced pressure, and purified by column chromatography on silica gel (petroleum ether/EtOAc = 5:1) to provide benzyl 4-(5-(5-(ethoxycarbonyl)-3-methyl- 1H -pyrazol-1-yl)pent-1-yn-1-yl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylate (9.5 g, 20.5 mmol, 45.3% yield) as a yellow solid. LCMS [M+H] ⁺ = 463.

Step 6: 4-(5-(5-(Ethoxycarbonyl)-3-methyl- 1H -pyrazol-1-yl)pentyl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid

A mixture of 4-(5-(5-(ethoxycarbonyl)-3-methyl- 1H -pyrazol-1-yl)pent-1-yn-1-yl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid benzyl ester (19.0 g, 41.10 mmol), 10%Pd/C (0.22 g, 2.05 mmol) and THF (500 mL) was stirred at room temperature under a hydrogen atmosphere (4 atm) for 2 days. Then, the reaction mixture was filtered and concentrated under reduced pressure. The obtained residue was recrystallized from EtOAc/petroleum ether (1:5, v/v) to provide 4-(5-(5-(ethoxycarbonyl)-3-methyl-pyrazol-1-yl)pentyl)-1-ethyl-3-methyl-pyrazole-5-carboxylic acid (10.5 g, 27.90 mmol, 67.9% yield). ¹ H-NMR (400 MHz, CDCl ₃ ) δ ppm 6.63 (s, 1 H), 4.57-4.48 (m, 4 H), 4.38-4.32 (m, 2 H), 2.74-2.62 (m, 2 H), 2.32 (s, 3 H), 2.23 (s, 3 H), 1.91-1.8 6 (m, 2 H), 1.59-1.54 (m, 2 H), 1.45-1.37 (m, 8 H). LCMS [M + H] ⁺ = 377.

Step 7: 4-4-(7-(5-Carboxy-3-methyl- 1H -pyrazol-1-yl)heptyl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid

2 M NaOH aqueous solution (60 mL, 119.5 mmol) was added to a suspension of 4-(5-(5-(ethoxycarbonyl)-3-methyl- 1H -pyrazol-1-yl)pentyl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid (9.0 g, 23.9 mmol) in MeOH (120 mL) and water (120 mL) stirred at room temperature. The reaction mixture was stirred at room temperature for 30 min. Then, the mixture was acidified to pH 4 by adding 6 M HCl solution, and a solid was precipitated from the reaction mixture afterwards. The solid was collected by filtration and dried under reduced pressure to provide 4-(5-(5-carboxyl-3-methyl- 1H -pyrazol-1-yl)pentyl)-1-ethyl-3-methyl- 1H -pyrazole-5-carboxylic acid (6.5 g, 18.7 mmol, 78.1% yield) as a white solid. ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ ppm 6.57 (s, 1 H), 4.40-4.34 (m, 4 H), 2.53 (t, J = 7.6 Hz, 2 H), 2.16 (s, 3 H), 2.09 (s, 3 H), 1.74-1.67 (m, 2 H), 1 .44-1.37 (m, 2 H), 1.27-1.16 (m, 5 H). LCMS [M + H] ⁺ = 349.

Step 8: 1-Allyl-2-amino-1H-benzo[d]imidazole-5-carboxamide, hydrobromide

To a solution of 4-fluoro-3-nitrobenzamide (10.0 g, 54.3 mmol) in DMF (60 mL) was added allylamine (36.6 mL, 489 mmol) dropwise at room temperature and the mixture was stirred for 5 min. After this time, K ₂ CO ₃ (15.01 g, 109 mmol) was added in one portion and the mixture was stirred at room temperature for 30 min. Then, the DMF was removed in vacuo. The residue was suspended in 500 mL of water and the resulting orange precipitate was filtered off, washed with water and dried in vacuo.

The above precipitate was dissolved in AcOH (600 mL) and the flask was placed in a 20°C water bath. Zinc (10.65 g, 163 mmol) was carefully added in small portions. The reaction was monitored by LCMS and additional zinc (approximately 3eq) was added in small portions as needed until the reduction was complete. After the reaction was complete as measured by LCMS, the solid was filtered off and the filtrate was concentrated in vacuo. The evaporation residue was taken up in DCM (500 mL) and EtOH (150 mL) and washed with 15% K ₂ CO ₃ aqueous solution (100 mL). The organic layer was separated, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo.

The above evaporation residue was dissolved in MeOH (200 mL), 5.0M cyanogen bromide (11.95 mL, 59.7 mmol) in CH ₃ CN was added quickly in one portion, and the mixture was stirred at room temperature for 18 h. After this time, the reaction mixture was concentrated in vacuo and then dissolved in MeOH (200 mL) again. A mixture of toluene (100 mL) and CH ₃ CN (100 mL) was added, and the resulting mixture was concentrated to dryness at 40° C. (0-1 mbar) and dried in vacuo for 16 h to provide 1-allyl-2-amino-1H-benzo[d]imidazole-5-carboxamide, hydrobromide (11.3 g, 38.0 mmol, 70.0% yield) as a dark purple powder. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.83 (s, 2 H), 8.07 (br. s.,1 H), 7.88 (d , J =1.00 Hz, 1 H), 7.82 (dd, J =8.41, 1.38 Hz, 1 H), 7.52 (d, J = 8.53 Hz, 1 H LCMS [M + H ] ⁺ = 216.9.

Step 9: 1-allyl-2-(1-(5-(5-((1-allyl-5-carbamoyl-1H-benzo[d]imidazol-2-yl)carbamoyl)-1-ethyl-3-methyl-1H-pyrazol-4-yl)pentyl)-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

A 5.0 mL ^Biotage® sealed tube was charged with 4-(5-(5-carboxy-3-methyl-1H-pyrazol-1-yl)pentyl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (634 mg, 1.820 mmol), 1-allyl-2-amino-1H-benzo[d]imidazole-5-carboxamide, hydrobromide (1352 mg, 4.55 mmol), HATU (1730 mg, 4.55 mmol) and NMP (13 mL). After stirring at room temperature for 1 minute, DIPEA (3.17 mL, 18.20 mmol) was added and the mixture was stirred at room temperature for 5 min and then heated at 140 °C in a microwave reactor for 1 h. After this time, 5.0 mL of water was added and the mixture was stirred at room temperature for 5 min. It was then poured into 250 mL of ice-cold water and stirred vigorously for 1 h. The resulting solid was filtered off, washed with water, dissolved from the filter using MeOH/DCM, concentrated in vacuo, and subjected to silica gel chromatography ( ^Biotage® UltraSNAP 100 g SiO _column : 0-40% MeOH/EtOAc) to afford 1-allyl-2-(1-(5-(5-((1-allyl-5-carbamoyl-1H-benzo[d]imidazol-2-yl)carbamoyl)-1-ethyl-3-methyl-1H-pyrazol-4-yl)pentyl)-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (840 mg, 1.128 mmol, 62% yield) as a pink solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.88 (s, 1 H), 12.81 (s, 1 H), 7.99 - 8.02 (m, 2 H), 7.97 (br. s., 2 H), 7.77 (ddd, J =8.34, 3.66, 1.39Hz, 2 H), 7 , 4.50 - 4.61 (m, 4 H), 2.73 (t, J =7.45 Hz, 2 H), 2.15 (s, 3 H), 2.08 (s, 3H), 1.71 - 1.85 (m, 2 H), 1.45 - 1.55 (m, 2 H), 1.27 - 1.34 (m, 5 H); LCMS [M + H] ⁺ = 745.7.

Step 10: 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,31-dodecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-1][1,3,6,15,17]pentaazacyclohexene-3,24-dicarboxamide

Four 20 mL ^Biotage® microwave sealed tubes were charged with a total of 1-allyl-2-(1-(5-(5-((1-allyl-5-carbamoyl-1H-benzo[d]imidazol-2-yl)carbamoyl)-1-ethyl-3-methyl-1H-pyrazol-4-yl)pentyl)-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (160 mg, 0.215 mmol), Hoveyda-Grubbs II catalyst (26.9 mg, 0.043 mmol) and freshly degassed 1,2-dichloroethane (DCE) (80 mL). The sealed tubes were heated at 100 °C in a microwave reactor for 4 h. After the mixture was cooled to room temperature, MeOH (1.0 mL) was added to each tube and the resulting clear solution was stirred at room temperature for 5 min. Potassium 2-isocyanatoacetate solution (15 mg in 1.5 mL MeOH) was added to each tube and the resulting mixture was stirred at room temperature for 5 min. The tubes were combined, concentrated in vacuo, and the evaporation residue was taken up in a minimum volume of DCM/MeOH and purified by silica gel chromatography ( ^Biotage® UltraSNAP 100 g _SiO2 column; 0-40% MeOH/EtOAc) to obtain the desired product (61 mg) as a light green solid containing a mixture of olefin isomers. The product was further purified ( ^Biotage® Ultra SNAP 25 g SiO2 _column ; 0-20% MeOH/DCM gradient) to afford 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,31-dodecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-1][1,3,6,15,17]pentaazacyclohexenedecene-3,24-dicarboxamide (54 mg, 0.075 mmol, 35% yield) as a 7:1 trans:cis mixture. Characterization of trans isomers: ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm12.87 (s, 1 H), 12.84 (s, 1 H), 7.98 (br. s., 4 H), 7.77 (dd, J =7.71, 3.16Hz, 2 H), 7.33 - 7.48 (m, 4 H), 6.55 (s, 1 H), 5.89 - 5.98 (m, 1 H), 5.66 -5.75 (m, 1 H), 4.90 (d, J =7.83 Hz, 4 H), 4.73 (t, J =6.95 Hz, 2 H), 4.47 (q, J =6.99 Hz, 2 H), 2.72 - 2.80 (m, 2 LCMS [M + H ] ⁺ =717.6.

Step 11: 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31-tetradecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-1][1,3,6,15,17]pentaazacyclohexenedecene-3,24-dicarboxamide

A round bottom flask was charged with 10% Pd/C (200 mg, 0.188 mmol) and purged with nitrogen. A solution of 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,31-dodecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-1][1,3,6,15,17]pentaazacyclohexene-3,24-dicarboxamide (100 mg, 0.140 mmol, 7:1 trans:cis mixture) in a mixture of MeOH (20.0 mL) and THF (20.0 mL) was added, the flask was purged with hydrogen, and the reaction mixture was stirred under hydrogen atmosphere (1 atm) for 23 h. The flask was then opened to air, stirred vigorously for 15 min and filtered, the Pd/C was washed with MeOH/THF, the filtrate was concentrated in vacuo and subjected to silica gel chromatography ( ^Biotage® Ultra SNAP 25 g _SiO2 column; 0-20% MeOH/DCM) to afford 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31-tetradecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-1][1,3,6,15,17]pentaazacyclohexenedecene-3,24-dicarboxamide (56 mg, 0.078 mmol, 4% by weight) as a light pink solid. 55.8% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.88 (br. s.,2 H), 8.02 (s, 4 H), 7.79 - 7.87 (m, 2 H), 7.67 (d, J =8.34 Hz, 1 H), 7.63 (d, J =8.34 Hz, 1 H), 7.37 ( br. s., 2 H), 6.57 (s, 1 H), 4.74 (t, J =6.57 Hz, 2 H), 4.48 (q, J =6.99 Hz, 2 H), 4.19 - 4.31 (m, 4 H), 2.78 - 2.86 (m, 2 H), 2.16 (s, 3 H), 2.08 ( s, 3 H), 1.91 (br. s., 4 H), 1.77 - 1.86 (m, 2 H), 1.44 -1.54 (m, 2 H), 1.35 - 1.42 (m, 2 H), 1.29 (t, J =7.07 Hz, 3 H); LCMS (m/z): 719.7 [M + H] ⁺ .

Step 12: (5aE,21E)-8-ethyl-5,10,18,22-tetramethyl-7,20-dioxo-5,7,8,11,12,13,14,15,20,22,28,29,30,31-tetradecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-l][1,3,6,15,17]pentaazacyclohexene-3,24-dicarboxamide

To a solution of 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31-tetradecahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4',3'-1][1,3,6,15,17]pentaazacyclohexene-3,24-dicarboxamide (85 mg, 0.118 mmol) in DMF (3 mL) was added iodomethane (0.015 mL iodomethane, 0.236 mmol, 83 uL stock solution of 180 uL iodomethane in 820 uL DMF). The reaction was stirred at room temperature for 3 h. The reaction was dry loaded onto silica gel and purified by silica gel chromatography (ISCO-Rf, 12 g column, 0%-30% gradient of MeOH/DCM) to afford a light yellow solid. The material was suspended in MeCN and concentrated to dryness under a stream of air over the weekend to afford (5aE,21E)-8-ethyl-5,10,18,22-tetramethyl-7,20-dioxo-5,7,8,11,12,13,14,15,20,22,28,29,30,31-tetrahydrobenzo[4,5]imidazo[1,2-a]benzo[4,5]imidazo[2,1-p]dipyrazolo[5,1-e:4′,3′-1][1,3,6,15,17]pentaazacyclohexeneicosadecadecene-3,24-dicarboxamide (58 mg, 0.077 mmol, 65.0% yield) as a white solid. ¹ H NMR (400MHz, methanol- d ₄ ) δ ppm 8.08 (dd, J =6.97, 1.39 Hz, 2 H), 7.87 - 7.99 (m, 2 H), 7.63 (d, J =8.62 Hz, 1 H), 7.51 (d, J =8.62 Hz, 1 H), 6.53 (s, 1 H), 4.63 (t, J =6.97 Hz, 2 H), 4.52 (q, J =7.18 Hz, 2 H), 4.20 - 4.32 (m, 4 H), 3.61 (s, 6H), 2.73 - 2.84 (m, 2 H), 2.26 (s, 3 H), 2.21 (s, 3 H), 1.87 (br. s., 6 H), 1.53 - 1.63 (m, 2 H), 1.30 - 1.40 (m, 5 H). LCMS m/z = 747 [M + H] ⁺ .

Example 4

(E)-1-((E)-4-((E)-5-Carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

Step 1: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (78 mg, 0.068 mmol) in DMF (3 mL) was added cesium carbonate (66.1 mg, 0.203 mmol) and 2.5 eq iodomethane (50 uL stock solution made from 220 uL iodomethane in 780 uL DMF). The reaction was stirred at room temperature for 4 h. A white precipitate formed in the yellow reaction mixture. Add other iodomethane (2.5 eq), and the reaction loses its yellow color immediately. Stir the reaction at room temperature over the weekend. Need other cesium carbonate (66 mg, 0.20 mmol) and iodomethane solution (2.5 eq) to drive the reaction to be complete. Add water and adopt DCM (3x) then adopt ~15% EtOH/DCM (2x) aqueous layer extraction. Adopt salt water washing to merge organic extract, dried over sodium sulfate and be concentrated to dryness. The resulting residue was dry loaded onto a 12 g silica gel column and eluted with a 0-20% gradient of MeOH/DCM to afford (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (20 mg, 0.022 mmol, 33.1% yield) as a white solid. LCMS m/z = 893 [M + H] ⁺ .

Step 2: (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

To a solution of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (20 mg, 0.022 mmol) in 1,4-dioxane (0.5 mL) was added 4 M HCl in dioxane (0.011 mL, 0.045 mmol). Additional 4 M HCl in dioxane was added as needed to drive deprotection. When the reaction was complete, the reaction was filtered and the filter cake washed with dioxane and dried in a vacuum oven at 45°C overnight. The resulting light yellow solid was believed to be (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (17 mg, 0.020 mmol, 87% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.01 - 8.13 (m, 3 H) 7.78 - 7.84 (m, 1H) 7.75 (s, 1 H) 7.35 - 7.52 (m, 4 H) 6.42 (s, 2 H), 5.88 - 5.97 (m, 1 H),5 .60 - 5.68 (m, 1 H), 4.88 (d, J =5.07 Hz, 2 H) 4.76 (d, J =5.58 Hz, 2 H) 4.36- 4.48 (m, 4 H) 4.11 (t, J =6.34 Hz, 2 H) 3.56 (s, 3 H) 3.53 (s, 3 H) 3.46 (s, 2 H) 2.12 (s, 6 H) 1.75 (d, J =6.08 Hz, 2 H) 1.21 (t, J =7.10 Hz, 6 H). LCMS m/z = 779 [M + H] ⁺ .

Example 5

(E)-1-((E)-4-((Z)-5-Carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride

Step 1: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((Z)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (50 mg, 0.058 mmol) in DMF (2 mL) was added cesium carbonate (75 mg, 0.231 mmol) and iodoethane (27.0 mg, 0.173 mmol). After 3 h, additional iodoethane (15 uL) was added and the reaction was stirred for 15 min. The reaction was partitioned between DCM and water. The aqueous layer was extracted with DCM/EtOH (3x). The combined organics were washed with brine, dried over sodium sulfate, dry loaded onto silica gel and purified by silica gel chromatography (12 g column, 0-20% MeOH/DCM gradient) to afford (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((Z)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (18 mg, 0.020 mmol, 33.8% yield). LCMS m/z = 921 [M+H] ⁺ .

Step 2: (E)-1-((E)-4-((Z)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

To a solution of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((Z)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (17 mg, 0.018 mmol) in 1,4-dioxane (923 µL) was added HCl in dioxane (27.7 µL, 0.111 mmol). After 1 h at room temperature, the reaction was filtered and the filter cake was washed with diethyl ether and dried in a vacuum oven at 55 °C overnight to provide (E)-1-((E)-4-((Z)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (14 mg, 0.015 mmol, 81% yield) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.03 - 8.14 (m, 3 H), 7.68 - 7.87 (m, 2 H), 7.42 - 7.52 (m, 4 H), 6.38 (s, 2 H), 5.80 - 5.97 (m, 1 H), 5.41 - 5.68 (m, 1 H), 4.81 - 4.94 (m, 2 H), 4.67 - 4.78 (m, 2 H), 4.34 - 4.45 (m, 4 H), 4.01 - 4.22 (m, 7 H), 3.32 - 3.51 (m, 2 H), 2.12 (s, 3 H), 2.10 (s, 3 H),1.68- 1.76 (m, 2 H), 1.11 - 1.32 (m, 12 H). LCMS m/z = 807 [M+H] ⁺ .

The compounds prepared by the above methods may exist as tautomers/isomers, for example (E)-1-((E)-4-((E)-5-carbamoyl-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-3-ethyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride

。

.

Example 6

(2E,2'E)-1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide), trifluoroacetate

Step 1: 4,4'-((2,2,3,3-tetrafluorobutane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide)

4-Fluoro-3-nitrobenzamide (3.59 g, 19.5 mmol) was added to 2,2,3,3-tetrafluorobutane-1,4-diamine (1.25 g, 7.81 mmol) and potassium carbonate (3.24 g, 23.4 mmol) in DMF (50 mL) at room temperature over 5 min, and the reaction was stirred overnight. The mixture was quenched with water, and the resulting solid was collected by filtration and ground with MeOH to provide the title compound (600 mg, 1.23 mmol, 16% yield) as a yellow solid. LCMS [M+H] ⁺ = 489.

Step 2: 4,4'-((2,2,3,3-tetrafluorobutane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide)

4,4'-((2,2,3,3-tetrafluorobutane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide) (1.15 g, 2.36 mmol) and palladium on _carbon (0.251 g, 2.36 mmol) in MeOH (100 mL) were stirred under H2 at 30 °C overnight. The reaction was filtered and the filtrate was concentrated to give the title compound (250 mg, 0.584 mmol, 25% yield). LCMS [M+H] ⁺ = 429.1.

Step 3: 1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-amino-1H-benzo[d]imidazole-5-carboxamide)

To 4,4'-((2,2,3,3-tetrafluorobutane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide) (20 mg, 0.047 mmol) in MeOH (1 mL) and water (2 mL) was added cyanogen bromide (29.7 mg, 0.280 mmol) and the reaction was stirred at 30°C overnight. The MeOH was removed in vacuo and the resulting solid was collected by filtration to give the title compound (15 mg, 0.031 mmol, 67% yield). LCMS [M+H] ⁺ = 479.0.

Step 4: 1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide)

To HATU (763 mg, 2.01 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (227 mg, 1.47 mmol) in DMF (20 mL) was added EDC (385 mg, 2.01 mmol), 1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-amino-1H-benzo[d]imidazole-5-carboxamide) (320 mg, 0.667 mmol) and DIEA (0.467 mL, 2.68 mmol) in one portion at room temperature. The reaction was heated to 70 °C for 12 h, concentrated and purified to give the title compound (8 mg, 0.01 mmol, 2% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 13.05 (s, 2 H), 8.01 (d, J =8.6 Hz, 4 H), 7.81 (d, J =8.2 Hz, 2 H), 7.53 (d, J =8.3 Hz, 2 H), 7.38 (s, 2 H), 6.73 (s, 2 H ), 5.32 (t, J =16.0 Hz, 4 H), 4.59 (dd, J =14.0, 6.9Hz, 4 H), 2.06 (s, 6 H), 1.33 (t, J =7.1 Hz, 6 H); LCMS [M+H] ⁺ = 751.1.

Step 5: (2E,2'E)-1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide), trifluoroacetate

To a 100 mL round bottom flask was added 1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (49 mg, 0.065 mmol) and DMF (0.653 mL). To the solution was added cesium carbonate (63.8 mg, 0.196 mmol) followed by iodomethane (10.20 µL, 0.163 mmol). The mixture was stirred at room temperature. After 30 minutes, more iodomethane (10 uL; 0.16 mmol) was added and the mixture was stirred at room temperature overnight (~14 hours). This clear crude mixture was directly injected into a reverse phase preparative HPLC system and purified (biphasic ISCO, 20-50% CH ₃ CN / H ₂ O gradient, TFA modifier). Pure fractions were combined and concentrated to afford (2E,2'E)-1,1'-(2,2,3,3-tetrafluorobutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide), trifluoroacetate salt (3 mg, 3.19 µmol, 4.89% yield) as an off-white semisolid. ¹ H NMR (400 MHz, methanol- d 4) δ ppm 1.34 (t, J =7.10 Hz, 6 H) 2.25 (s, 6 H) 3.70 (s, 6 H) 4.60 (q, J =7.10 Hz, 4 H) 5.19 (t, J =15.33 Hz, 4 H) 6.66 (s, 2 H) 7.60 (d, J =8.36 Hz, 2 H) 7.96 (dd, J =8.36, 1.52Hz, 2 H) 8.11 (d, J =1.27 Hz, 2 H). LCMS m/z = 779 [M+H] ⁺ .

Example 7

1-((E)-4-((E)-5-Carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate

Step 1: (E)-1-((E)-4-((4-Carbamoyl-2-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

4-Fluoro-3-nitrobenzamide (86 mg, 0.467 mmol), (E)-1-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 3 hydrochloride (name used in PU66420P: (Z)-1-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 3 hydrochloride (250 mg, 0.467 mmol) and DIPEA (0.245 4-[4-[4-[4-[4-(4-[4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4- ⁽ 4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-(4-

Step 2: (E)-1-((E)-4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-((E)-4-((4-carbamoyl-2-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (105 mg, 0.178 mmol) in acetic acid (0.500 mL) and MeOH (0.5 mL) was added 1 wt % Pt and 2 wt % Vanadium on activated carbon - 50-70% wet powder (34.7 mg, 1.781 µmol, Strem, 78-1536). The flask was evacuated and purged with nitrogen, and this was repeated two more times. The flask was evacuated and flushed with a hydrogen balloon, and stirred at room temperature under a hydrogen atmosphere. Due to incomplete conversion after 5 h, MeOH is used to filter the reaction mixture through a small diatomite plug. The reaction mixture is concentrated under vacuum and stored in a refrigerator. The crude material is dissolved in acetic acid (0.500mL) and MeOH (0.5 mL) again, and 1 wt% Pt and 2 wt% vanadium V-50-70% wetted powder (34.7 mg, 1.781 μmol) on activated carbon are added. The flask is emptied and purged with nitrogen, which is repeated twice more. The flask is emptied and rinsed with a hydrogen balloon, and stirred at room temperature under a hydrogen atmosphere. After 2 h, MeOH is used to filter the reaction mixture through a small diatomite plug, then concentrated under vacuum to obtain the title compound (163 mg, 0.148 mmol, 82% yield) as a red oil. LCMS m/z=280 [M+2H/2] ⁺ .

Step 3: 1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate

To (E)-1-((E)-4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (100 mg, 0.179 mmol) in DMF (1 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (0.491 mL, 0.197 mmol, 0.4 M in dioxane) at 0°C and stirred at 0°C for 1 h. After 1 h, EDC (51.4 mg, 0.268 mmol) and triethylamine (0.075 mL, 0.536 mmol) were added and the reaction was stirred at 40°C for 3 h and at room temperature overnight. The reaction is diluted with 1.5 mL DMSO and the title compound is purified by mass-guided preparative HPLC (alkaline modifier). Pure fractions are collected and organic matter is removed under vacuum. Then, the compound is extracted with 2 x 25 mL DCM, and the organic layer is washed with 10 mL salt water. Volatiles are removed under vacuum to provide the title compound (by LCMS～80% purity). The compound is diluted with 2.0 mL DMSO and purified again by mass-guided preparative HPLC (TFA modifier). Pure fractions are collected and solvent is removed under vacuum to provide the title compound (15 mg, 0.016 mmol, 8.8% yield) as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 1.31 - 1.38 (m, 6 H), 2.24 - 2.26 (m, 6 H), 3.71 (s, 3 H), 3.88 (s, 3 H), 4.50 (q, J =7.10 Hz, 2 H), 4.60 (q, J =7.10 Hz, 2 H), 4.88 (d, J =5.83 Hz, 2 H), 5.11 (d, J =5.07 Hz, 2 H), 5.78 (dt, J =15.40, 5.73 Hz, 1 H), 5.95 - 6.03 (m, 1 H), 6.63 (s, 1 H), 6.65(s, 1 H ), 7.36 (d, J =8.36 Hz, 1 H), 7.51 (d, J =1.27 Hz, 1 H), 7.74 (dd, J =8.36, 1.52 Hz, 1 H), 7.79 (d, J =1.01 Hz, 1 H), 8.00 (d, J =1.52 Hz, 1 H). LCMSm/z = 721 [M + H] ⁺ .

Example 8

1-((E)-4-((E)-5-Carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate

Step 1: (E)-1-((E)-4-((4-Carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

4-Chloro-3-methoxy-5-nitrobenzamide (108 mg, 0.467 mmol), (E)-1-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 3 hydrochloride (250 mg, 0.467 mmol) and DIPEA (0.245 mL, 1.402 mmol) were suspended in isopropanol (2 mL) and heated at 120 °C in a sealed vial. After 22 h, the reaction was diluted with 25 mL EtOAc and washed with 2 x 25 mL water, 25 mL saturated sodium bicarbonate solution and 25 mL brine. The aqueous layer was extracted with 25 mL EtOAc. The organic layer is collected and concentrated under vacuum to provide a crude product as an orange solid. The crude product is dissolved in 12 mL DMSO, filtered and directly purified by mass-guided preparative HPLC (high pH modifier, multiple injections). Pure fractions are merged, organic matter is removed under vacuum, and the title compound is extracted by aqueous solvent using 2 x 50 mL DCM. Volatiles are removed under vacuum to provide the title compound (53 mg, 0.086 mmol, 18.3% yield) as an orange solid. LCMS m/z = 620 [M + H] ⁺ .

Step 2: (E)-1-((E)-4-((2-amino-4-carbamoyl-6-methoxyphenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To (E)-1-((E)-4-((4-carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (name used in PU66420P: (Z)-1-((E)-4-((4-carbamoyl-2-methoxy-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (50 mg, 0.081 mmol)) in acetic acid (0.500mL) and MeOH (0.5 mL) suspension was added to 1 wt% Pt and 2 wt% vanadium-50-70% wet powder on activated carbon (15.74 mg, 0.807 μmol, Strem, 78-1536). The vial was evacuated and purged with nitrogen, and it was repeated twice more. The vial was evacuated and rinsed with a hydrogen balloon, and then stirred at room temperature under a hydrogen atmosphere. After 4 h, the reaction mixture was filtered through a small diatomaceous earth plug using MeOH, and then concentrated under vacuum to provide a crude product mixture (52 mg) as a light red solid. LCMS m/z = 590 [M + H] ⁺ .

Step 3: 1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate

To (E)-1-((E)-4-((2-amino-4-carbamoyl-6-methoxyphenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (60 mg, 0.102 mmol) in DMF (1 mL) at 0 °C was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (0.280 mL, 0.112 mmol) as a 0.4 M solution in dioxane and stirred at 0 °C for 1 h. After 1 h, EDC (29.3 mg, 0.153 mmol) and triethylamine (0.043 mL, 0.305 mmol) were added and the reaction was stirred at 40 °C for 2 h, then at room temperature for 18 h. The reaction mixture was diluted with 1.5 mL DMSO and purified on mass-directed preparative HPLC (high pH modifier). A second preparative HPLC purification step (using TFA-modifier) was required to provide the pure title compound (5.0 mg, 5.11 mmol, 5.0% yield) as an off-white solid. ¹ HNMR (400 MHz, DMSO- d ₆ ) δ ppm 1.21 (t, J =7.10 Hz, 3 H) 1.26 (t, J =7.10 Hz, 3H) 2.10 (s, 3 H) 2.12 (s, 3 H) 3.50 (s, 3 H) 3.75 (s, 3 H) 3.80 (s, 3 H) 4.43(q, J =7.18 Hz, 2 H) 4.53 (q, J =7.01 Hz, 2 H) 4.82 - 4.91 (m, 4 H) 5.76 - 5.80(m, 2 H) 6.36 (s, 1 H) 6.52 (s, 1 H) 7.33 (d, J =1.01 Hz, 1 H) 7.38 (br. s., 1H) 7.43 (d, J =0.76 Hz, 1 H) 7.48 (br. s, 1 H) 7.65 (d, J =1.27 Hz, 1 H) 7.72 (d, J =1.01 Hz, 1 H) 7.99 (br. s., 1 H) 8.05 ( br. s, 1 H) 12.85 (s, 1 H). LCMSm/z = 751 [M + H] ⁺ .

Example 9

1-((E)-4-((E)-5-Carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

Step 1: ((E)-1-((E)-4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-chloro-5-nitrobenzamide (182 mg, 0.467 mmol), (E)-1-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 3 hydrochloride (250 mg, 0.467 mmol) and DIPEA (0.245 mL, 1.402 mmol) were suspended in isopropanol (2 mL) and heated in a sealed vial at 120° C. After 22 h, the reaction was diluted with 25 mL EtOAc and washed with 2 x 25 mL water, 25 mL saturated sodium bicarbonate solution and 25 mL brine. The aqueous layer was back extracted with 25 mL EtOAc. The organic layer was collected and concentrated under vacuum. The crude product was dissolved in 6 mL DMSO, filtered and purified by mass-oriented preparative HPLC (high pH modifier). Pure fractions were combined, organic matter was removed under vacuum, and 2 x 50 mL DCM was used to extract the compound by aqueous solvent. Solvent evaporation provided the title compound (80 mg, 0.103 mmol, 22% yield) as an orange solid. LCMS m/z = 778 [M +H] ⁺ .

Step 2: (E)-1-((E)-4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

(E)-1-((E)-4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (name used in PU66420P: A suspension of (Z)-1-((E)-4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide) (80 mg, 0.103 mmol) in acetic acid (0.500 mL) and MeOH (0.5 mL) was added 1 wt % Pt and 2 wt % Vanadium on activated carbon - 50-70% wet powder (20.06 mg, 1.028 µmol, Strem 78-1536). The vial was evacuated and purged with nitrogen and this was repeated two more times. The vial was evacuated and flushed with a hydrogen balloon and stirred at room temperature under a hydrogen atmosphere. After 4 h, the reaction mixture was filtered through a small celite plug using MeOH, then concentrated and dried under vacuum to provide the title compound (77 mg, 0.072 mmol, 70% yield). LCMS m/z = 748 [M + H] ⁺ .

Step 3: 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To (E)-1-((E)-4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (80 mg, 0.107 mmol) in DMF (1 mL) at 0 °C was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (0.294 mL, 0.118 mmol, 0.4 M) in dioxane and stirred at 0 °C for 1 h. EDC (30.8 mg, 0.160 mmol) and triethylamine (0.045 mL, 0.321 mmol) were added, and the reaction was stirred at 40 ° C for 2 h and kept at room temperature for 18 h. The reaction was diluted with 1.5 mL DMSO and purified using mass-oriented preparative HPLC (high pH modifier). Pure fractions were collected and organic matter was removed under vacuum. Then, the compound was extracted with 2 x 25 mL DCM, and the organic layer was washed with 10 mL brine. Volatiles were removed under vacuum to provide the title compound (25 mg, 0.027 mmol, 26% yield) as an off-white solid. LCMSm/z = 909 [M+H] ⁺ .

Step 4: 1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

To 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (25 mg, 0.027 mmol) in MeOH (1 mL) was added hydrochloric acid in dioxane (0.069 mL, 0.275 mmol, 4 M) and the reaction was stirred at room temperature. After 10 min, the volatiles were removed under vacuum to afford the title compound as a white solid (22 mg, 0.025 mmol, 92% yield). ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 1.29 - 1.36 (m, 3 H), 1.45 (t, J =6.72 Hz, 3 H), 1.85 - 1.91 (m, 2 H), 2.29 (s, 3 H), 2.36 (s, 3 H), 3.64 (t, J =5.4 5 Hz, 2 H), 3.89 (s, 3 H), 3.91 (s, 3 H), 4.18 (t, J =5.32 Hz, 2 H), 4.38 (q, J =6.51 Hz, 2 H), 4.69 (q, J =6.80 Hz, 2 H), 5.19 (br. s., 2 H), 5.30 (br . s., 2 H), 5.85 - 6.06 (m, 2 H), 6.80 (s, 1 H), 6.99 (s, 1 H), 7.43 (s, 1 H), 7.61 (s, 1 H), 7.74 (s, 1 H), 8.00 (s, 1 H). LCMS m/z = 795 [M + H] ⁺ .

Example 10

(E)-1-((E)-4-((E)-5-Carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate

Into a 20-mL vial was placed (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (200 mg, 0.235 mmol) and DMF (2.35 mL). To this solution was added cesium carbonate (230 mg, 0.706 mmol) and iodomethane (37 µL, 0.588 mmol). The solution was stirred at room temperature for 15 min. DMF (2 mL) and water (2 mL) were added directly to the vial. This mixture was directly purified using mass-directed preparative HPLC (15-55% gradient of MeCN/water with _NH4OH as modifier). The corresponding fractions were combined and concentrated. The concentrated mixture was further purified by mass-directed preparative HPLC (5-35% gradient of MeCN/water with TFA as modifier). The corresponding fractions were combined and concentrated to afford (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2 trifluoroacetate salt (3 mg, 2.69 umol, 1.14% yield) as a clear oil. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm9.94 (br. s., 2 H), 8.09 (br. s., 2 H), 7.81 (s, 1 H), 7.78 (s, 1 H), 7.53 (br. s., 2 H), 7.42 (d, J =6.08 Hz, 2 H), 6.42 (m, 2 H), 5.51 - 5.85 (m, 2 H), 4.72 - 4.99 (m, 4 H), 4.42 (q, J =6.84 Hz, 4 H), 4.05 (t, J =5.58 Hz, 2 H), 3.95 (d, J =11.66 Hz, 2 H), 3.69 (s, 3H), 3.62 (t, J =11.91 Hz, 2 H), 3.56 (s, 3 H), 3.54 (s, 3 H), 3.34 (d, J =11.91 Hz, 2 H), 3.19 (d, J =7.10 Hz, 2 H), 3.02 (br. s., 2 H), 2.12 (s, 6 H), 1.94 ( m, 2 H), 1.21 (m, 6 H). LCMS ( m/z ):878.7 [M + H] ⁺ .

Embodiment 11

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-(4-methyl-4-morpholino)propoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate, trifluoroacetate

Into a 20-mL vial was placed (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (200 mg, 0.235 mmol) and DMF (2.35 mL). To this solution was added cesium carbonate (230 mg, 0.706 mmol) and iodomethane (37 µL, 0.588 mmol). The solution was stirred at room temperature for 15 min. DMF (2 mL) and water (2 mL) were added directly to the vial. This mixture was directly purified using mass-directed preparative HPLC (15-55% gradient of MeCN/water with _NH4OH as modifier). The corresponding fractions were combined and concentrated. The concentrated mixture was purified by reverse phase preparative HPLC (5-35% gradient of MeCN/water with TFA as modifier) using an acidic modifier. The corresponding fractions were combined and concentrated to provide the title compound (6 mg, 4.81 umol, 2.04% yield) as a clear oil. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.08 (br. s.,2 H), 7.81 (s, 2 H), 7.77 (s, 1 H), 7.53 (br. s., 3 H), 7.39-7.40 (m, 3 H), 6.41 (s, 1 H), 6.40 (s, 1 H), 5.60-5.75 (m, 2 H), 4.74 - 4.97 (m, 4 H), 4.35-4.51 (m, 4 H), 4.02 (t, J =5.32 Hz, 2 H), 3.82-3.98 (m, 4 H), 3.67 (s, 3 H), 3.50-3.54 (m, 8H), 3.08 (s, 3 H), 2.12 (s, 6 H), 1.95-2.05 (m., 2 H), 1.24-1.20 (m, 6 H). LCMS ( m/z ): 892.7 [M] ⁺ .

Example 12

(2E,2'E)-1,1'-(Pentane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide)

Step 1: 4,4'-(pentane-1,4-diylbis(azanediyl))bis(3-nitrobenzamide)

Into a reaction flask was placed pentane-1,4-diamine, 2-hydrochloride (1 g, 5.71 mmol) and isopropanol (9.52 mL). To this solution was added 4-fluoro-3-nitrobenzamide (1.052 g, 5.71 mmol) followed by DIPEA (4.49 mL, 25.7 mmol). The flask was capped and the reaction was heated to 105 °C. After 4 hours, more 4-fluoro-3-nitrobenzamide (1.052 g, 5.71 mmol) and isopropanol (10 mL) were added. The mixture was stirred at 105 °C overnight (~14 h). The formed precipitate was filtered off and rinsed twice with isopropanol (5 mL each). 4,4'-(Pentane-1,4-diylbis(azanediyl))bis(3-nitrobenzamide) (2.6 g, 5.80 mmol, 100% yield) was obtained as a yellow solid. LCMS ( m/z ): 431.3 [M + H] ⁺ .

Step 2: 4,4'-(pentane-1,4-diylbis(azanediyl))bis(3-aminobenzamide)

To a 100-mL round bottom flask was added 4,4'-(pentane-1,4-diylbis(azanediyl))bis(3-nitrobenzamide) (500 mg, 1.162 mmol) and MeOH (11.6 mL). To this solution was added ammonium chloride (249 mg, 4.65 mmol) and 5.5 mL of saturated aqueous ammonium chloride solution. To this solution was added zinc (759 mg, 11.62 mmol). The heterogeneous mixture was stirred at room temperature for 15 min. The mixture was filtered and the collected solids were rinsed with MeOH (10 mL). Celite was added to the combined filtrates and the crude product was purified by flash chromatography (dry loading technique, 12 g SiO2 _cartridge , 2-40% MeOH/DCM as eluent, with _NH4OH as modifier). The corresponding fractions were combined and concentrated. 4,4'-(pentane-1,4-diylbis(azanediyl))bis(3-aminobenzamide) (368 mg, 0.944 mmol, 81% yield) was obtained as a colorless oil. LCMS ( m/z ): 371.2 [M + H] ⁺ .

Step 3: 1,1'-(Pentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide)

To a 100-mL round bottom flask was added 4,4'-(pentane-1,4-diylbis(azanediyl))bis(3-aminobenzamide) (368 mg, 0.993 mmol) and DMF (9.9 mL). The solution was cooled to 0°C. After stirring at 0°C for 5 min, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (3 mL of a ~0.4 M solution in dioxane, ~1.2 mmol) was added. After 15 min, more 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (1 mL of a ~0.4 M solution in dioxane, ~0.4 mmol) was added. The reaction was stirred at 0°C for an additional 15 min. Then, EDC (476 mg, 2.483 mmol) and triethylamine (0.692 mL, 4.97 mmol) were added. The reaction mixture was allowed to warm to room temperature and stirred overnight (~14 h). The reaction mixture was poured into 4:1 water/saturated aqueous ammonium chloride solution (25 mL). The product was extracted with ethyl acetate (3 x15 mL). The combined organic phase was washed with water (20 mL), brine (20 mL) and dried over magnesium sulfate. The crude material was concentrated and purified by flash chromatography (24 g SiO ₂ cartridges, 2-40% MeOH/DCM as eluent, containing NH ₄ OH as a modifier). The corresponding fractions were combined and concentrated. 1,1'-(pentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (303 mg, 0.429 mmol, 43.1% yield) was obtained as a white powder. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.88 (br. s, 1 H), 12.80 (br. s, 1 H), 7.95 - 7.99 (m, 4 H), 7.70 - 7.73 (m, 2 H), 7.61 (d, J =8.62 Hz, 1 H), 7.34 - 7.42 (m, 3 H), 6.61 (m, 2 H), 5.20 (br. s., 1 H), 4.53- 4.60 (m, 4 H), 4.32 (br. s, 1 H), 4.11 - 4.16 (m, 1 H), 2.34 (br. s, 1 H), 2.10 (s, 3 H) , 2.09 (s, 3 H), 1.92 (m, 1 H), 1.70 (m, 1 H), 1.61 (m, 1 H), 1.52 (d, J =6.84 Hz, 3 H), 1.28 - 1.34 (m, 6 H); LCMS (m/z): 693.6 [M + H] ⁺ .

Step 4: (2E,2'E)-1,1'-(Pentane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide)

Into a 10-mL vial was placed 1,1'-(pentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (50 mg, 0.072 mmol) and DMF (1.4 mL). To the heterogeneous solution was added cesium carbonate (70.5 mg, 0.217 mmol) followed by iodomethane (0.011 mL, 0.180 mmol). The vial was capped and the mixture was stirred at room temperature overnight (~14 h). The mixture was diluted with DMSO (1 mL) and (1 mL) to form a clear homogeneous solution. This solution was directly purified by reverse phase preparative HPLC (biphasic ISCO system, 5-35% gradient of MeCN/water with 0.1% NH ₄ OH modifier). The corresponding fractions were combined and concentrated. The product was lyophilized with MeCN and water (~30 mL). (2E,2'E)-1,1'-(pentane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide) was obtained as a white fluffy solid (25 mg, 0.035 mmol, 48.1% yield). ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 7.87 (d, J =2.28 Hz, 2 H), 7.82 (dd, J =8.36, 1.52 Hz, 1 H), 7.73 - 7.80 (m, 1 H), 7.54 (d, J =8.36 Hz, 1H), 7.41 (d, J =8.62 Hz, 1 H), 6.63 (s, 2 H), 4.85 (m, 1 H), 4.63 (q, J =7.18Hz, 4 H), 4.25 - 4.41 (m, 1 H), 4.06 - 4.20 (m, 1 H), 3.57 (s, 3 H), 3.55 (s, 3 H), 2.25 (s, 6 H), 2.23 (m, 1 H), 1.81 - 2.03 (m, 2 H), 1.68 (m., 1 H), 1.58 (d, J =6.84 Hz, 3 H), 1.39 (t, J =7.10 Hz, 6 H). LCMS (m/z): 721.6 [M + H] ⁺ .

Example 13

(2E,2'E)-1,1'-(4-methylpentane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide)

Step 1: 4,4'-((4-methylpentane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide)

Into a 24-mL vial was placed 4-methylpentane-1,4-diamine (0.49 g, 4.22 mmol) and isopropanol (14.0 mL). To this solution was added 4-fluoro-3-nitrobenzamide (1.63 g, 8.85 mmol) followed by DIPEA (2.58 ml, 14.76 mmol). The vial was capped and the heterogeneous solution was stirred at 105 °C overnight (~14 h). The precipitate was filtered off and rinsed with isopropanol (2 x 5 mL). 4,4'-((4-methylpentane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide) (1.46 g, 3.02 mmol, 71.7% yield, 92% purity) was obtained as an orange solid. LCMS ( m/z ): 445.3 [M + H] ⁺ .

Step 2: 4,4'-((4-methylpentane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide)

To a 100-mL round bottom flask was added 4,4'-((4-methylpentane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide) (500 mg, 1.035 mmol) and MeOH (15 mL). To this solution was added ammonium chloride (1107 mg, 20.70 mmol), 10 mL of saturated aqueous ammonium chloride, and zinc (677 mg, 10.35 mmol). The heterogeneous mixture was stirred at room temperature. After 20 min, more zinc (350 mg, 5.35 mmol) and ammonium chloride (600 mg, 11.22 mmol) were added. After stirring for a total of 90 min at room temperature, the mixture was filtered. The remaining solid was rinsed with MeOH (20 mL). Celite was added to the combined filtrate and the crude product was purified by silica gel chromatography (dry loading technique, 12 g SiO2 _cartridge , 2-40% gradient of MeOH/DCM with _NH4OH as modifier). The corresponding fractions were combined and concentrated. 4,4'-((4-methylpentane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide) (247 mg, 0.610 mmol, 59.0% yield) was obtained as a white film. LCMS ( m/z ): 385.4 [M+H] ⁺ .

Step 3: 1,1'-(4-methylpentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide)

To a 100-mL round-bottom flask was added 4,4'-((4-methylpentane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide) (247 mg, 0.642 mmol) and DMF (6.42 mL). This solution was cooled to 0°C. After stirring at 0°C for 5 min, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~0.4 M in dioxane, 2.5 mL; ~1.0 mmol) was added as a solution. After 20 min, more 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~0.4 M in dioxane, 0.45 mL; 0.18 mmol) was added to ensure complete thiourea formation. After stirring at 0 °C for an additional 20 min, EDC (308 mg, 1.606 mmol) was added followed by triethylamine (0.448 mL, 3.21 mmol). The reaction mixture was allowed to warm to room temperature and stirred overnight (~14 h). The reaction was poured into a beaker containing 25 mL of 3:1 water/saturated aqueous ammonium chloride solution and stirred for 10 min. The resulting white precipitate was filtered off and rinsed with water (3x 5 mL). The solid was dried in a vacuum oven at 50 °C for 6 h. 1,1'-(4-methylpentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (341 mg, 0.473 mmol, 73.6% yield) was obtained as an off-white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.93 (m, 3 H), 7.84 (s, 1 H), 7.73 (d, J =8.87 Hz, 1 H), 7.67 (d, J =8.36 Hz, 1 H), 7.60 (d, J =8.62 Hz, 1 H), 7.24 - 7 .44 (m, 2 H), 6.40 (s, 1 H), 6.25 (br. s., 1 H), 4.39- 4.64 (m, 4 H), 4.11 (t, J =6.46 Hz, 2 H), 2.44 (br. s., 2 H), 2.10 (s, 3 H), 2.07 (s, 3 H), 1.90 (br. s., 6 H), 1.68 (br. s., 2 H), 1.31 (t, J =7.10 Hz, 3H), 1.24 (t, J =7.10 Hz, 3 H). LCMS ( m/z ): 707.6 [M + H] ⁺ .

Step 4: (2E,2'E)-1,1'-(4-methylpentane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide)

Into a 10-mL vial was placed 1,1'-(4-methylpentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (49 mg, 0.069 mmol) and DMF (0.693 mL). To this solution was added cesium carbonate (67.8 mg, 0.208 mmol) followed by iodomethane (10.84 µL, 0.173 mmol). The vial was capped and the mixture was stirred at room temperature overnight (~14 h). The mixture was diluted with DMSO (1 mL) and water (1 mL) to form a clear homogeneous solution. This solution was injected directly and purified by reverse phase preparative HPLC (biphasic ISCO system, 5-35% gradient of MeCN/water with 0.1% NH ₄ OH modifier). The corresponding fractions were combined and concentrated. (2E,2'E)-1,1'-(4-methylpentane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide) was obtained as a clear oil (15.8 mg, 0.022 mmol, 31.0% yield). ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 7.64 - 7.85 (m, 5 H), 7.34 (d, J =8.36 Hz, 1 H), 6.65 (s, 1 H), 6.57 (s, 1 H), 4.64 (m, 4 H), 4.20 (t, J =5.96Hz, 2 H), 3.54 (s, 3 H), 3.40 (s, 3 H), 2.24 - 2.31 (m, 5 H), 2.23 (s, 3 H), 1.82 - 1.95 (m, 8 H), 1.41 (m, 6 H). LCMS ( m/z ): 735.4 [M + H] ⁺ .

Embodiment 14

(E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2-methylpentan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-N,3-dimethyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a 10-mL vial was added 1,1'-(4-methylpentane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (synthetic intermediate of Example 13) (100 mg, 0.141 mmol) and DMF (1.415 mL). To this solution was added cesium carbonate (138 mg, 0.424 mmol) followed by iodomethane (0.044 mL, 0.707 mmol). The vial was capped and the mixture was stirred at room temperature overnight (~14 h). This mixture was directly purified by reverse phase preparative HPLC (biphasic ISCO system, gradient of MeCN/water with 0.1% _NH4OH modifier). The corresponding fractions were combined and concentrated. (E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2-methylpentan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-N,3-dimethyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (7.1 mg, 0.0087 mmol, 6.16% yield) was obtained as a white film. ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 7.56 - 7.85 (m, 5 H), 7.25 - 7.37 (m, 1 H), 6.66 (s, 1 H), 6.58 (s, 1 H), 4.54 - 4.76 (m, 4 H), 4.20 (t, J =5.96 Hz , 2 H), 3.55 (d, J =5.58 Hz, 3H), 3.40 (s, 3 H), 3.02 (s, 3 H), 2.26 (s, 3 H), 2.26 (m, 1 H), 2.23 (s, 3H), 2.23 (m, 1 H), 1.87 (s, 6 H), 1.87 (m, 2 H), 1.42 (m, 6 H). LCMS ( m/z ):749.4 [M + H] ⁺ .

Embodiment 15

(E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (Diastereomer 1)

Step 1: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(-5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Into a 10-mL vial was placed 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (25 mg, 0.028 mmol, e.g. Intermediate 14B, second eluting diastereomer) and DMF (559 µL). To this heterogeneous solution was added cesium carbonate (27.3 mg, 0.084 mmol) followed by iodomethane (4.37 µL, 0.070 mmol). The vial was capped and the mixture was stirred at room temperature overnight (~14 h). The mixture was diluted with DMSO (1 mL) and water (1 mL) to form a clear homogeneous solution. This solution was injected directly and purified by reverse phase preparative HPLC (biphasic ISCO system, 3-35% gradient of MeCN/water with 0.1% _NH4OH modifier) and the corresponding fractions were combined and concentrated to afford (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(-5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (18 mg, 0.019 mmol, 66.3% yield) as a white solid. LCMS: 923.4 [M + H] ⁺ , 1.30 min retention time (Acquity UPLC CSH C18, 1.7 um, 50 mm x 2.1 mm column; 3-95% gradient over 1.5 min, MeCN/10 mM ammonium bicarbonate in water adjusted to pH 10 with 25% ammonium hydroxide solution).

Step 2: (E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

Into a 20-mL vial was placed (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(-5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (25 mg, 0.027 mmol) and MeOH (2 mL). To this solution was added HCl (3 M in CPME, 300 µL, 0.900 mmol). The reaction mixture was stirred at room temperature overnight. The volatiles were removed under _a N ₂ blow down unit.The crude product was purified by reverse phase preparative HPLC (biphasic ISCO system, 20-50% gradient of MeCN/water with 0.1% NH ₄ OH modifier). The corresponding fractions were combined and concentrated to afford (E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (5.5 mg, 6.46 µmol, 23.85% yield) as a glassy white solid. ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 8.01 (s, 1.17) 7.88 (br. s., 3.21) 7.49 - 7.73 (m, 8.82) 7.42 (br. s., 2.89) 7.29 (br. s., 0.86) 7.20 (s, 2 .93) 6.70 (s, 3.36) 6.64 (br. s., 2.95) 4.98 - 5.11(m, 4.53) 4.81 (br. s., 3.36) 4.55 - 4.75 (m, 14.25) 4.29 - 4.55 (m, 3.26)4.08 (m, s., 4.09) 2.19 - 2.36 (m, 21.72) 2.00 - 2.18 (m, 11.47) 1.76 - 2.00 (m, 13.31) 1.54 (m,23.37) 1.35 - 1.50 (m, 24.00); LCMS: 809.5 [M + H] ⁺ , 0.74 min retention time (AcquityUPLC CSH C18 50 mm x 2.1 mm column, 1.7 um; 5-95% gradient over 1.5 min; MeCN/water with 0.1% TFA modifier).

Example 16

(E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (Diastereomer 2)

Step 1: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Into a 10-mL vial was placed 7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (25 mg, 0.028 mmol, e.g. Intermediate 14A, first eluting diastereomer) and DMF (559 µL). To this heterogeneous solution was added cesium carbonate (27.3 mg, 0.084 mmol) and iodomethane (4.37 µl, 0.070 mmol). The vial was capped and the mixture was stirred at room temperature overnight (~14 h). The mixture was diluted with DMSO (1 mL) and water (1 mL) to form a clear homogeneous solution. This solution was directly injected and purified by reverse phase preparative HPLC (biphasic ISCO system, 5-35% gradient of MeCN/water with 0.1% NH ₄ OH modifier). The corresponding fractions were combined and concentrated to afford (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (15 mg, 0.014 mmol, 49.5 % yield, 85 % purity) as a white solid. LCMS: 923.5 [M + H] ⁺ ,1.24 min retention time (Acquity UPLC CSH C18, 1.7 um, 50 mm x 2.1 mm column; 3-95% gradient over 1.5 min, MeCN/10 mM ammonium bicarbonate in water, adjusted to pH 10 with 25% ammonium hydroxide solution).

Step 2: (E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

Into a 20-mL vial was placed (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(-5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (15 mg, 0.016 mmol) and MeOH (2 mL). To this solution was added HCl (3 M in CPME, 300 µL, 0.900 mmol). The reaction mixture was stirred at room temperature overnight. Volatiles were removed using a stream of nitrogen. The residue was purified by reverse phase preparative HPLC (biphasic ISCO system; 20-50% gradient of MeCN/water with 0.1% _NH4OH modifier). The corresponding fractions were combined and concentrated to afford racemic (E)-1-(5-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)hexan-2-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (5 mg, 5.87 μmol, 36.1% yield) as a glassy white solid. ¹ H NMR (400 MHz, methanol- d ₄ ) δ ppm 8.07 (m, 2.61), 7.86 (m, 2.78), 7.58 - 7.81 (m, 4.92), 7.52 (s, 2.38), 6.48 - 6.73 (m, 4.98), 5.47 (br. s. , 1.43), 4.75 (br. s., 2.43), 4.49 - 4.69 (m, 10.33), 4.26 - 4.40 (m, 4.06), 4.21 (br. s., 1.08), 3.50 - 3.76 (m, 20.52), 2.68 (s, 1.17) , 2.49 (br.s., 0.84), 2.15 - 2.41 (m, 18.94), 1.94 - 2.07 (m, 2.86), 1.78 - 1.94 (m,3.63), 1.70 (m., 3.14), 1.46 - 1.63 (m, 16.31), 1.26 - 1.42 (m, 17.00). LCMS: 809.5 [M + H] ⁺ , 0.71 min retention time (Acquity UPLC CSH C18, 1.7 um, 50 mm x 2.1 mm column; 5-95% gradient over 1.5 min; MeCN/water with 0.1% TFA modifier).

Embodiment 17

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide, 2-trifluoroacetate

Step 1: 3-(3-(Benzyloxy)propoxy)-4-chloro-5-nitrobenzamide

To a stirred suspension of 4-chloro-3-hydroxy-5-nitrobenzamide (30 g, 139 mmol) and potassium carbonate (57.4 g, 416 mmol) in DMF (200 mL) was added a solution of ((3-bromopropoxy)methyl)benzene (47.6 g, 208 mmol) dropwise at room temperature under nitrogen over 1 min. The reaction mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature and quenched by adding 200 mL of water. Then, the aqueous phase was extracted with DCM (3x100 mL). The combined organic layers were washed with water (4x200 mL) and brine (200 mL), dried, and concentrated under vacuum. The crude product was purified by silica gel chromatography (100 g column, 1:2 petroleum ether/EtOAc). Appropriate fractions were combined and concentrated to provide 3-(3-(benzyloxy)propoxy)-4-chloro-5-nitrobenzamide (33 g, 90 mmol, 65.3% yield) as a yellow solid. LCMS ( m/z ): 365 [M + H] ⁺ .

Step 2: 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-(3-(benzyloxy)propoxy)-5-nitrobenzamide

A suspension of 2,2,3,3-tetrafluorobutane-1,4-diamine (4 g, 24.98 mmol), 3-(3-(benzyloxy)propoxy)-4-chloro-5-nitrobenzamide (4.56 g, 12.49 mmol) and DIPEA (6 mL, 34.4 mmol) in isopropanol (18 mL) was stirred overnight at 135 °C in a sealed tube. When cooled, the volatiles were removed in vacuo. The crude product was purified by silica gel chromatography (20 g column, 60-100% gradient of EtOAc/petroleum ether). The appropriate fractions were combined and concentrated to provide 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-(3-(benzyloxy)propoxy)-5-nitrobenzamide (3 g, 6.14 mmol, 24.6% yield) as a reddish brown oil. LCMS ( m/z ): 489 [M + H] ⁺ .

Step 3: 3-(3-(Benzyloxy)propoxy)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-nitrobenzamide

A suspension of 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-(3-(benzyloxy)propoxy)-5-nitrobenzamide (3 g, 6.14 mmol), 4-fluoro-3-nitrobenzamide (1.696 g, 9.21 mmol) and potassium carbonate (1.698 g, 12.28 mmol) in DMF (30 mL) was stirred overnight at 60 °C under nitrogen. When cooled, water (50 mL) was added and the aqueous phase was extracted with EtOAc (2x100 mL). The combined organic layers were washed with water (3x200 mL) and brine (200 mL), dried and concentrated under vacuum. The crude product was purified by silica gel chromatography (10 g column, 30-100% gradient of EtOAc/petroleum ether). Appropriate fractions were combined and concentrated to afford 3-(3-(benzyloxy)propoxy)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-nitrobenzamide (2 g, 3.06 mmol, 49.9% yield) as a yellow solid. LCMS ( m/z ): 653 [M+H] ⁺ .

Step 4: 3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-(3-(benzyloxy)propoxy)benzamide

To a stirred suspension of 3-(3-(benzyloxy)propoxy)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-nitrobenzamide (1.9 g, 2.91 mmol) in acetic acid (20 mL) under nitrogen was added solid zinc (1.904 g, 29.1 mmol) in one portion. The reaction mixture was stirred at 25 °C for 2 h. The reaction solution was then filtered and the filtrate was concentrated under vacuum to provide 3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-(3-(benzyloxy)propoxy)benzamide (1.5 g, 2.53 mmol, 87% yield) as a brown solid. LCMS ( m/z ): 593 [M+H] ⁺ .

Step 5: 2-amino-1-(4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-(3-(benzyloxy)propoxy)-1H-benzo[d]imidazole-5-carboxamide

To a solution of 3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-(3-(benzyloxy)propoxy)benzamide (2.1 g, 3.54 mmol) in MeOH (20 mL) was added cyanogen bromide (1.126 g, 10.63 mmol). The reaction mixture was stirred at 25 °C for 16 h. The mixture was diluted with diethyl ether (30 mL). The mixture was filtered and the filter cake was washed with diethyl ether. The filtrate was concentrated under reduced pressure to afford 2-amino-1-(4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-(3-(benzyloxy)propoxy)-1H-benzo[d]imidazole-5-carboxamide (1.8 g, 2.381 mmol, 67.2% yield, -85% purity) as a grey solid. The product was used directly without further purification. LCMS ( m/z ): 643 [M+H] ⁺ .

Step 6: 7-(3-(Benzyloxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide

To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (0.864 g, 5.60 mmol), 2-amino-1-(4-(2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-(3-(benzyloxy)propoxy)-1H-benzo[d]imidazole-5-carboxamide (1.8 g, 2.80 mmol) and DIPEA (1.957 mL, 11.20 mmol) in DMF (20 mL) was added HATU (2.66 g, 7.00 mmol). The reaction mixture was stirred at 60 °C for 16 h. The mixture was poured into water. The precipitate was collected by filtration, washed with water, MeCN, and diethyl ether, and then dried under vacuum to afford 7-(3-(benzyloxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (1.1 g, 1.05 mmol, 37.3% yield, -87% purity) as a light brown solid. The product was used without further purification. LCMS ( m/z ): 915 [M+H] ⁺ .

Step 7: 1-(4-(5-Carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazol-5-carboxamide

To a solution of 7-(3-(benzyloxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (1.1 g, 1.2 mmol) in MeOH (30 mL) and NMP (10.0 mL) was added palladium on carbon (1.279 g). The reaction was hydrogenated at 60 °C using an H-cube system (4 atm hydrogen) for 72 h. The mixture was diluted with DMF (20 mL). The catalyst was then removed by filtration and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (Gemini-C18, 5µ silica gel, 21x150 mm column, 10-60% gradient of MeCN/water with 0.1% TFA modifier). Pure fractions were combined and evaporated to dryness to afford 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazole-5-carboxamide (75 mg, 0.086 mmol, 7.19% yield) as a pink solid. ¹ H NMR (400 MHz, DMSO) δ 13.06-13.04 (m, 2 H), 8.06-8.03 (m, 3 H), 7.84 (d, J = 8.4 Hz, 1 H), 7.70 (s, 1 H), 7.53 (d, J =8.4 Hz, 1 H), 7.46 (s, 1 H), 7.40 (s, 2 H), 6.66-6.65 (m, 2 H), 5.27-2.20 (m,4 H), 4.62-4.50 (m, 5 H), 4.26 (t, J = 6.1 Hz, 2 H), 3.55 (t, J = 5.7 Hz, 2H), 2.06 (s, 6 H), 1.98-1.82 (m, 2 H), 1.33 (t, J = 7.0 Hz, 6 H). LCMS ( m/z ):824.4 [M + H] ⁺ .

Step 8: (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate

Into an 8-mL vial was placed 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazole-5-carboxamide (25 mg, 0.030 mmol), cesium carbonate (49.4 mg, 0.152 mmol), and DMF (1 mL). To this solution was added iodomethane (4.26 µL, 0.068 mmol). The vial was capped and the mixture was stirred at room temperature overnight (~14 h). The sample was diluted with more DMF and directly purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of MeCN/water with 0.1% TFA modifier). The corresponding fractions were combined and concentrated in vacuo to afford (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, 2-trifluoroacetate salt (2 mg, 1.702 µmol, 5.62% yield) as a clear solid. ¹ H NMR (DMSO- d ₆ , 600 MHz): δ ppm 8.09 (s, 2 H), 8.05 (br. s., 1 H), 7.89 (dd, J =8.4, 1.2 Hz, 1 H), 7.75 (s, 1 H), 7.57 (d, J =8.3 Hz, 1 H), 7.51 (s , 1 H), 7.48 (br. s., 1 H), 7.46 (br. s., 1H), 6.52 (s, 1 H), 6.51 (s, 1 H), 5.15-5.30 (m, 4 H), 4.51-4.48 (m, 4 H), 4.22 (br. t., J =6.5 Hz, 2 H ), 3.60 (s, 3 H), 3.57 (s, 3 H), 3.51 (br. t., J =6.0 Hz, 2 H), 2.15 (s, 3 H), 2.12 (s, 3 H), 1.84 (br. t., J =6.3 Hz, 2 H), 1.24-1.26 (m, 6 H). LCMS (m/z): 853.4 [M + H] ⁺ .

Embodiment 18

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: (E)-4-((4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide

To a solution of (E)-2-(4-amino-2,3-dimethylbut-2-en-1-yl)isoindoline-1,3-dione (5.7 g, 23.33 mmol) and 4-fluoro-3-nitrobenzamide (3.9 g, 21.18 mmol) in DMSO (65 mL) was added potassium carbonate (6.44 g, 46.6 mmol). The reaction was stirred at room temperature for 3 h. The reaction mixture was poured into a flask containing 300 mL of rapidly stirring water. The mixture was stirred for 5 min, filtered and the filtered solid was rinsed sequentially with water, diethyl ether (2x) and ethyl acetate (2x). The solid was collected, stirred in hexanes (30 mL), filtered and dried to provide (E)-4-((4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (6.77 g, 16.6 mmol, 78% yield) as a bright yellow solid. LCMS ( m/z ): 409.2 [M+H] ⁺ .

Step 2: (E)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide

To a suspension of (E)-4-((4-(1,3-dioxoisoindolin-2-yl)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (6.4 g, 15.67 mmol) in EtOH (100 mL) was added hydrazine monohydrate (0.84 mL, 17.2 mmol). After 10 min, EtOH (50 ml) was added to facilitate stirring. The reaction mixture was heated at 80 °C for 20 h. The reaction was filtered while still warm and the desired product was found in both the solid and the filtrate. The solid and filtrate were combined, concentrated to dryness and used crude in the next reaction.

To a bright yellow suspension of (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (4.65 g, 10.53 mmol) and 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (3.30 g, 9.80 mmol) in 1-butanol (100 mL) was added sodium bicarbonate (2.47 g, 29.4 mmol). The mixture was heated at 120 °C. After 18 h, the reaction mixture was cooled to room temperature, stirred for 15 min and the solid was filtered and rinsed with n-butanol (2x25 mL). One portion of saturated NaHCO ₃ solution was diluted with one portion of water. The solid was washed with diluted NaHCO ₃ solution (2x25 mL), water (1x25 mL), diluted NaHCO ₃ solution (50 mL) and water (30 mL). The solid was collected in a round bottom flask and stirred in a diluted bicarb solution (100 mL) at room temperature for 2 h. The solid was filtered, washed with water and dried in a vacuum oven to obtain (E)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (3.62 g, 6.3 mmol, 63.8% yield) as an orange solid. LCMS ( m/z ): 579.3 [M + H] ⁺ .

Step 3: (E)-3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-((4-methoxybenzyl)oxy)benzamide

To a suspension of (E)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (3.46 g, 5.98 mmol) in MeOH (25 mL) and acetic acid (20 mL) was added 1% Pt and 2% V on activated carbon (50-70% wet powder, 1.167 g, 0.060 mmol). The flask was stirred at room temperature under a hydrogen atmosphere (hydrogen balloon) for 6 h. MeOH (10 mL) was used to rinse the solids off the sides of the flask and stirring was continued for an additional 16 h. After removal of the hydrogen, the reaction was filtered through celite, rinsed with MeOH and concentrated to provide a thick orange oil. DCM (15 mL) was added, and with stirring, the resulting mixture was treated with saturated NaHCO solution ( ₁ mL portion until bubbling stopped and the aqueous layer was alkaline). The liquid was decanted and the remaining solid was distributed between 3:1 CHCl ₃ :EtOH and saline. The organic layer was dried over sodium sulfate and concentrated to provide a light brown foam. The foam was ground with MeOH to provide (E)-3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,3-dimethylbut-2-ene-1-yl)amino)-5-((4-methoxybenzyl)oxy)benzamide (1.4 g, 2.70 mmol, 45.1% yield). LCMS ( m/z ): 519.4 [M+H] ⁺ .

Step 4: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-5-carboxamide

To an ice-cold solution of (E)-3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-((4-methoxybenzyl)oxy)benzamide (1.40 g, 2.70 mmol) in DMF (18 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~1 M in dioxane, 5.40 mL, 5.40 mmol) dropwise over 2 min. After 15 min, EDC (1.29 g, 6.75 mmol) and TEA (1.881 mL, 13.50 mmol) were added. The reaction was allowed to warm to room temperature and heated at 40 °C for 22 h. The reaction was cooled to room temperature and poured into a rapidly stirring 3:1 water:saturated aqueous NH ₄ Cl solution (100 mL). A fine solid formed immediately and the mixture was stirred for an additional 10 min. The solid was filtered, washed twice with water (50 mL) and dried in a vacuum oven at 50 °C overnight to provide (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (2.20 g, 2.48 mmol, 92% yield) as a light brown solid. ¹ H NMR (DMSO-d ₆ ) δ: 12.97 (d, J=3.3 Hz, 2 H), 8.08 (s, 1 H), 8.03 (br. s., 1H), 7.96 (br. s., 1 H), 7.68-7.75 (m, 2 H), 7.50 (s, 1 H), 7.40 (d . 96 (s, 2 H), 4.88(br. s., 2 H), 4.53-4.62 (m, 4 H), 3.59 (s, 3 H), 2.10 (s, 3 H), 2.05 (s, 3H), 1.66 (s, 3 H), 1.32 (t, J=7.0 Hz, 6 H), 1.17 (s, 3 H). LCMS ( m/z ): 841.5[M + H] ⁺ .

Step 5: (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (1.30 g, 1.546 mmol) in DMF (12 mL) was added cesium carbonate (1.511 g, 4.64 mmol) and iodomethane (0.242 mL, 3.86 mmol). The reaction mixture was stirred at room temperature for 1 h. Water (120 mL) was slowly added to the reaction mixture. The mixture was stirred vigorously for 30 min. The resulting solid was collected on a filter and dried. The crude product was purified by silica gel chromatography (80 g column, 30-80% gradient of (3:1 EA:EtOH with 1% ammonium hydroxide)/hexanes) to provide the title compound (1.01 g, 1.16 mmol, 74% yield) as a light yellow solid. ¹ H NMR (DMSO-d ₆ ) δ: 8.15 (s, 1 H), 8.10 (br. s., 1 H), 8.05 (br. s., 1 H), 7.77-7.82 (m, 2 H), 7.62 (s, 1 H), 7.52 (br. s., 1 H), 7.49 (br. s., 1 H), 7.22 (d, J=8.6 Hz, 1 H), 7.17 (d, J=8.6 Hz, 2 H), 6.68 (d, J=8.9 Hz, 2 H), 6.41 (s, 1 H), 6.38 (s, 1 H), 5.09 (s, 2 H), 4.96 (s, 2H), 4.77 ( s, 2 H), 4.48 (q, J=7.0 Hz, 4 H), 3.58-3.63 (m, 6 H), 3.56 (s, 3H), 2.07-2.11 (m, 6 H), 1.55 (s, 3 H), 1.22-1.28 (m, 6 H), 1.16 (s, 3 H). LCMS ( m/z ): 869.6 [M + H] ⁺ .

Embodiment 19

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a suspension of (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (930 mg, 1.070 mmol) in DCM (15 mL) was added HCl (4 M in dioxane, 1.34 mL, 5.35 mmol). The reaction mixture thickened. DCM (10 mL) and MeOH (2 mL) were added to obtain a homogeneous solution. The reaction was stirred at room temperature for 16 h. Because the reaction was not complete, the reaction mixture was concentrated and the solid residue was suspended in DCM (15 mL) and TFA (0.412 mL, 5.35 mmol). In 30 min, deprotection was complete. The reaction mixture was concentrated and the residue was distributed between 3:1 CHCl ₃ :EtOH and saturated NaHCO ₃ solution. The solid present was filtered and dried to provide the title compound (427 mg, 0.57 mmol). The remaining organic phase was concentrated and the residue was purified by silica gel chromatography (12 g silica gel column, 1-8% gradient of MeOH/DCM) to provide the title compound (130 mg, 0.17 mmol) as an additional amount of off-white solid. ¹ H NMR (DMSO-d ₆ ) δ ppm 10.68 (s, 1 H), 8.10 (s, 1H), 8.01 (br. s., 1 H), 7.96 (br. s., 1 H), 7.80 (d, J=8.4 Hz, 1 H), 7.62 (s,1 H), 7.46 (br. s., 1 H), 7.38 (br. s., 1 H), 7.32 (s, 1 H), 7.22 (d, J=8.4Hz, 1 H), 6.47 (s, 1 H), 6.33 (s, 1 H), 5.07 (s, 2 H), 4.83 (s, 2 H), 4.44-4.56 (m, 4 H) , 3.58 (s, 3 H), 3.53 (s, 3 H), 2.14 (s, 3 H), 2.07 (s, 3 H), 1.65 (s, 3 H), 1.48 (s, 3 H), 1.31-1.24 (m, 6 H). LCMS ( m/z ): 749.5 [M + H] ⁺ .

Embodiment 20

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide

To (E)-2,3-dimethylbut-2-ene-1,4-diamine, 2-hydrochloride (4.4 g, 23.52 mmol) and ethanol (81 ml) was added a solution of potassium carbonate (6.76 g, 48.9 mmol) in water (81 ml). Once all solids were dissolved, 4-fluoro-3-nitrobenzamide (3.0 g, 16.29 mmol) was added to the purple-brown solution in one portion at room temperature. The reaction mixture was stirred at room temperature for 105 min, then heated at 50 °C for 90 min and filtered. The filtrate was acidified with 6N HCl. The aqueous layer was washed with DCM (2x). The combined organics were extracted with water (1x). The combined aqueous layers were basified with 6N NaOH and saturated sodium bicarbonate and extracted with a 3:1 chloroform/ethanol mixture (3x). The combined organic extracts were dried over sodium sulfate, filtered and concentrated to dryness to afford (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (1.8 g, 5.30 mmol, 32.5 % yield) as a brownish yellow residue. LCMS ( m/z ): 279.1 [M+H] ⁺ .

Step 2: (E)-3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-((4-((4-carbamoyl-2-

(nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-nitrobenzamide

To a suspension of crude (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (715 mg, 2.312 mmol) in 1-butanol (10.9 mL) was added DIE (1.14 mL, 6.56 mmol). The mixture was stirred for 10 min and then 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-chloro-5-nitrobenzamide (850 mg, 2.186 mmol) was added. The reaction mixture was heated at 120 °C overnight. The reaction was cooled to room temperature and an orange solid precipitated. The mixture was filtered and the filter cake was washed with ethyl acetate to afford crude (E)-3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-nitrobenzamide (786 mg, 1.022 mmol, 46.8 % yield) as a bright orange solid, which still contained residual n-BuOH but was supported on it as is. LCMS ( m/z ): 631.3 [M+H] ⁺ .

Step 3: (E)-3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-(3-((tert-butyldimethylsilyl)oxy)propoxy)benzamide

To a mixture of (E)-3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-nitrobenzamide (1.82 g, 2.453 mmol) in methanol (53.3 ml) was added ammonium chloride (2.62 g, 49.1 mmol) followed by zinc (3.21 g, 49.1 mmol). The heterogeneous mixture was stirred at room temperature for 10 min. The reaction mixture was filtered and the filter cake was washed with methanol. The filtrate was dry loaded onto silica gel and purified by silica gel chromatography (80 g column, 0%-20% methanol, DCM with 0.1% NH ₄ OH modifier). The desired fractions were concentrated to dryness to afford (E)-3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-(3-((tert-butyldimethylsilyl)oxy)propoxy)benzamide (503 mg, 0.881 mmol, 35.9% yield) as a white solid. LCMS ( m/z ): 571.5 [M+H] ⁺ .

Step 4: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide

To a solution of (E)-3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-5-(3-((tert-butyldimethylsilyl)oxy)propoxy)benzamide (500 mg, 0.876 mmol) in DMF (8.8 mL) at 0°C was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~0.4 M, 4.82 mL, 1.93 mmol) in dioxane dropwise and portionwise. After 15 min, additional isothiocyanate (~0.4 M, 400 µL) was added and the reaction mixture was stirred for 10 min. The reaction was treated with EDC (420 mg, 2.190 mmol) and TEA (610 µl, 4.38 mmol) and stirred at room temperature over the weekend. The reaction mixture was poured into 4:1 water/saturated ammonium chloride (200 mL) and the resulting suspension was filtered. The filter cake was dried under a stream of nitrogen to provide (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (614 mg, 0.687 mmol, 78 % yield). ¹ HNMR (400 MHz, DMSO- d ₆ ) δ ppm 12.92 (d, J =11.66 Hz, 2 H), 7.99 - 8.08 (m, 2H), 7.90 (br. s., 1 H), 7.70 (s, 1 H), 7.65 (d, J =8.11 Hz, 1 H), 7.3 1 - 7.43(m, 3 H), 7.16 (d, J =8.36 Hz, 1 H), 6.60 (s, 1 H), 6.42 (s, 1 H), 5.14 (br.s., 2 H), 4.98 (br. s., 2 H), 4.48 - 4.66 (m, 4 H), 4.14 (t, J =5.58 Hz, 2 H), 3.65 (t, J =5.96 Hz, 2 H), 2.02 - 2.14 (m, 6 H), 1.61 - 1.74 (m, 6 H), 1.33 (t, 7.13 Hz, 3 H), 1.32 (t, 7.13 Hz, 3 H), 0.80 (s, 11 H), -0. 03 (s, 6 H). LCMS ( m/z ): 893.4 [M + H] ⁺ .

Step 5: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

A mixture of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (0.245 g, 0.274 mmol), cesium carbonate (0.268 g, 0.823 mmol) and iodomethane (0.043 mL, 0.686 mmol) in DMF (2 mL) was stirred at room temperature for 18 h. The reaction mixture was partitioned between EtOAc (20 mL) and brine (20 mL). The aqueous layer was extracted with EtOAc (2 x 10 mL).The combined EtOAc layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (RediSepRf High Performance Gold 40 g HP silica column, 40-80% gradient of (3:1 ethanol/ethyl acetate)/hexanes with 2% NH ₄ OH modifier) to afford (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (0.040 g, 0.033 mmol, 11.9% yield). LCMS ( m/z ): 921.7 [M + H] ⁺ .

Step 6: (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a mixture of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (0.030 g, 0.033 mmol) in MeOH (0.5 mL) was added HCl in dioxane (4 M, 0.163 mL, 0.651 mmol) and stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo. The crude product was purified by silica gel chromatography (RediSepRf High Performance Gold 40 gHP silica column, 40-90% gradient of (3:1 ethanol/ethyl acetate)/hexanes with 2% _NH4OH modifier) to afford (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (0.007 g, 7.72 µmol, 23.7% yield). ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 8.12 (s, 2H), 8.03 (br s, 1 H), 7.81 (d, J =8.66 Hz, 1 H), 7.78 (s, 1 H), 7.50 (s, 1 H), 7.47 (s, 1 H), 7.45 (br s, 1 H) , 7.24 (d, J =8.85 Hz, 1 H), 6.68 (br s, 1 H), 6.45 (s, 1 H), 6.34 (s, 1 H), 5.06 (br s, 2 H), 4.85 (s, 2 H), 4.49-4.53 (m, 2 H), 4.49 (br s, 2 H), 4.19 (br t, J =6.36 Hz, 2 H), 3.58 (s, 3 H), 3.56 (s,3 H), 3.44 (br t, J =5.99 Hz, 2 H), 2.10 (s, 3 H), 2.07 (s, 3 H), 1.72 (quint, J =6.27 Hz, 2 H), 1.61 (s, 3 H ), 1.48 (s, 3 H), 1.27 (t, J =7.10 Hz, 3 H), 1.24 (t, J =7.09 Hz, 3 H). LCMS (m/z): 807.6 [M + H] ⁺ .

Table 1 shows Examples 21-28, which can be prepared according to the following exemplified method:

Embodiment 29

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: (E)-4-((4-((2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide

To a bright yellow/orange suspension of (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide, hydrochloride (2.4 g, 7.62 mmol) and 2-fluoro-1-((4-methoxybenzyl)oxy)-3-nitrobenzene (2.114 g, 7.62 mmol) in DMF (20 mL) was added TEA (3.19 mL, 22.87 mmol). The thick mixture was stirred at room temperature for 2 h and then heated at 50 °C for 18 h. The reaction mixture was slowly added to rapidly stirring water (350 mL). The solid precipitated as a clumping mass. Sonication (30 min) and stirring for 1 h afforded a free flowing solid. The solid was filtered, rinsed with water and diethyl ether (3 x 30 mL), and dried to afford (E)-4-((4-((2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (3.52, 6.25 mmol, 82% yield) as a bright orange solid. LCMS ( m/z ): 536.2 [M+H] ⁺ .

Step 2: (E)-3-Amino-4-((4-((2-amino-6-((4-methoxybenzyl)oxy)phenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide

To a suspension of (E)-4-((4-((2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (3.52 g, 6.57 mmol) in methanol (15 mL) and acetic acid (10 mL) was added 1% Pt and 2% V on activated carbon - 50-70% wet powder (1.282 g, 0.066 mmol). The atmosphere of the flask was exchanged with nitrogen and then with hydrogen (balloon). After stirring at room temperature for 20 h, the atmosphere was exchanged with nitrogen. As the reaction was not complete, the reaction mixture was stirred under hydrogen atmosphere for an additional 5 h and then nitrogen was reintroduced. The reaction mixture was passed through celite and rinsed with 10% methanol/DCM. Concentration in vacuo afforded a thick orange oil. The oil was taken up in DCM (100 mL) and saturated aqueous sodium bicarbonate solution was added until bubbling ceased. The separation and concentration of the organic layer provide brown foam.Filterable solid is obtained by ethyl acetate containing a small amount of DCM and MeOH.The resulting solid is filtered and dried to provide (E)-3-amino-4-((4-((2-amino-6-((4-methoxybenzyl)oxy)phenyl)amino)-2,3-dimethylbut-2-ene-1-yl)amino)benzamide (1.95 g, 3.90 mmol, 59.3% yield) as a light brown solid.LCMS ( m/z ): 476.3 [M+H] ⁺ .

Step 3: (E)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-1H-benzo[d]imidazole-5-carboxamide

To a light brown solution of (E)-3-amino-4-((4-((2-amino-6-((4-methoxybenzyl)oxy)phenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide (1.93 g, 3.86 mmol) in DMF (20 mL) cooled in an ice/water bath was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~1 M in dioxane, 7.71 mL, 7.71 mmol) dropwise rapidly (over ~1 min). The reaction mixture was stirred for 25 min. EDC (1.848 g, 9.64 mmol) and TEA (2.69 mL, 19.28 mmol) were added and the reaction was warmed to room temperature and stirred for 18 h. The reaction mixture was poured into a rapidly stirred 1:1 saturated _NH4Cl aqueous solution:water (100 mL) to provide a fine precipitate. The precipitate was washed with water (2 x 15 mL), triturated twice with ethyl acetate (20 mL), and dried to provide (E)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-1-yl)-2,3-dimethylbut-2-ene-1-yl)-1H-benzo[d]imidazole-5-carboxamide (2.32 g, 2.83 mmol, 73% yield) as a brown solid. LCMS ( m/z ): 798.4[M+H] ⁺ .

Step 4: (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a solution of (E)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-1H-benzo[d]imidazole-5-carboxamide (1.09 g, 1.366 mmol) in DMF (15 mL) was added cesium carbonate (1.335 g, 4.10 mmol) and iodomethane (0.214 mL, 3.42 mmol). The reaction mixture was stirred at room temperature for 5 h. Additional iodomethane (0.060 mL, 0.956 mmol) was added and the mixture was stirred for an additional 18 h. Additional iodomethane (0.060 mL, 0.956 mmol) and cesium carbonate (1.335 g, 4.10 mmol) were added and the mixture was heated at 50°C for 4 h. The mixture was diluted with water (30 mL) and a sticky solid precipitated out. Vigorous stirring provided a filterable solid which was then collected on a filter and rinsed with water. Silica gel chromatography (40 g silica gel, 10-90% gradient of [3:1 EtOAc:EtOH]/heptane) afforded (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (430 mg, 0.509 mmol, 37% yield) as a brown foam. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.14 (s, 1 H), 8.01 (br. s., 1 H), 7.78 (dd, J =8.4, 1.4 Hz, 1 H), 7.45 (br. s., 1 H), 7.25-7.32 (m, 1 H), 7.13-7.2 3 (m, 4 H), 7.08 (d, J =8.0Hz, 1 H), 6.71 (d, J =8.5 Hz, 2 H), 6.43 (s, 1 H), 6.37 (s, 1 H), 5.03 (s, 2H), 4.97 (s, 2 H), 4.78 (s, 2 H), 4.45-4.5 3 (m, 4 H), 3.63 (s, 3 H), 3.60 (s, 3 H), 3.54 (s, 3 H), 2.10 (s, 3 H), 2.09 (s, 3 H), 1.56 (s, 3 H), 1.19-1.31 (m, 6 H), 1.18 (s, 3 H). LCMS ( m/z ): 826.5 [M + H] ⁺ .

Embodiment 30

(E)-1-((E)-4-((E)-7-(Benzyloxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a suspension of (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (45 mg, 0.055 mmol) in DMF (1 mL) was added (bromomethyl)benzene (7.26 µL, 0.061 mmol) followed by potassium carbonate (9.97 mg, 0.072 mmol). The reaction mixture was initially stirred at 50 °C for 3 h and then at room temperature for 16 h. The mixture was diluted with water. The aqueous layer was extracted several times with 3:1 CHCl ₃ :EtOH. The solvent was evaporated in vacuo and the residue was purified by mass-guided HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 30-85% gradient of MeCN/water with 0.1% TFA modifier). A few drops of saturated sodium bicarbonate solution were added to each clean fraction. ACN was removed using a stream of nitrogen. The suspended solid was filtered, rinsed with water, and dried to provide pure (E)-1-((E)-4-((E)-7-(benzyloxy)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (30 mg, 0.037 mmol, 67%) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.12 (s, 1 H), 8.01 (br. s., 1 H), 7.76 (dd, J=8.4, 1.4 Hz, 1 H), 7.44 (br. s., 1 H), 7.12-7.32 (m, 8 H), 7.07 (d, J =7.8 Hz, 1 H), 6.47 (s, 1H), 6.38 (s, 1 H), 5.13 (s, 2 H), 4.99 (s, 2 H), 4.78 (s, 2 H), 4.45-4.55 (m, 4 H), 3.59 (s, 3 H), 3.56 (s, 3 H), 2.1 3 (s, 3 H), 2.10 (s, 3H), 1.59 (s, 3H), 1.19-1.31 (m, 6H), 1.19 (s, 3H). LCMS ( m/z ): 796.6 [M + H] ⁺ .

Embodiment 31

(E)-1-((E)-4-((E)-4-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

Step 1: tert-Butyl (E)-(4-((3-bromo-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)carbamate

To tert-butyl (E)-(4-amino-2,3-dimethylbut-2-en-1-yl)carbamate (0.5 g, 2.33 mmol) in ethanol (11.67 mL) was added 1-bromo-3-fluoro-2-nitrobenzene (0.529 g, 2.33 mmol) and DIEA (1.22 mL, 7.00 mmol). The mixture was stirred at 80 °C for 18 h. The mixture was partitioned between ethyl acetate (50 mL) and brine (20 mL). The aqueous layer was extracted with EtOAc (2 x 10 mL). The combined EtOAc layers were dried over magnesium sulfate, filtered and concentrated. The residue was purified by silica gel chromatography (40 g silica gel, gradient of 10-20% ethyl acetate/hexanes) to afford tert-butyl (E)-(4-((3-bromo-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)carbamate (0.740 g, 1.78 mmol, 77% yield). LCMS ( m/z ): 414.1 [M + H] ⁺ .

Step 2: (E)-4-((4-((3-bromo-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide

To tert-butyl (E)-(4-((3-bromo-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)carbamate (0.44 g, 1.06 mmol) in methanol (5 mL) was added 4 M HCl in dioxane (1.06 mL, 4.25 mmol). The mixture was stirred at room temperature for 3 h. Additional 4 M HCl in dioxane (1.0 mL, 4.0 mmol) was added and stirred for another 3 hours. The reaction mixture was concentrated in vacuo to remove HCl and solvent. To this residue was added 4-fluoro-3-nitrobenzamide (0.214 g, 1.16 mmol), DIEA (0.927 mL, 5.31 mmol) and 1-butanol (15 ml) and the mixture was stirred at 110°C for 16 h. The mixture was partitioned between ethyl acetate (50 mL) and brine (20 mL). The aqueous layer was extracted with EtOAc (2 x 10 mL). The combined EtOAc layers were dried over magnesium sulfate, filtered and concentrated. Concentration provided a precipitate, which was filtered and dried to provide (E)-4-((4-((3-bromo-2-nitrophenyl)amino)-2,3-dimethylbut-2-ene-1-yl)amino)-3-nitrobenzamide (0.40 g, 0.836 mmol, 79% yield) as a solid. LCMS ( m/z ): 478.1 [M + H] ⁺ .

Step 3: (E)-3-Amino-4-((4-((2-amino-3-bromophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide

To (E)-4-((4-((3-bromo-2-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide (0.050 g, 0.105 mmol) in methanol (1.6 mL) was added 28-30% ammonium hydroxide solution (0.102 mL, 2.61 mmol) and sodium dithionite solution (0.214 g, 1.045 mmol, in 0.8 mL water). The mixture was stirred at room temperature for 2 h. The reaction mixture was partitioned between ethyl acetate (20 mL) and brine (20 mL). The aqueous layer was extracted with EtOAc (2 x 10 mL). The combined EtOAc layers were dried over sodium sulfate, filtered and concentrated. Purification by silica gel chromatography (12 g silica gel, 10-40% gradient of (3:1 ethanol/ethyl acetate)/heptane using 2% _NH4OH modifier) provided (E)-3-amino-4-((4-((2-amino-3-bromophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide (0.030 g, 0.070 mmol, 67.2% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 7.42 (br.s., 1 H), 7.01-7.15 (m, 2 H), 6.70 (d, J =7.10 Hz, 2 H), 6.38-6.47 (m, 1 H), 6.32 (dd, J =7.86, 14.95 Hz, 2 H), 5.01 (t, J =5.07 Hz, 1 H), 4.93 (t, J =5.07Hz, 1 H), 4.74 (s, 2 H), 4.63 (s, 2 H), 3.76 (d, J =4.82 Hz, 2 H), 3.71 (d, J =5.07 Hz, 2 H), 1.78 (br. s., 3 H), 1.77 (br. s., 3 H).

Step 4: (E)-1-(4-(4-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide

To (E)-3-amino-4-((4-((2-amino-3-bromophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide (0.030 g, 0.072 mmol) in DMF (0.72 mL) at 0°C was added a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (1 M in dioxane, 0.158 mL, 0.158 mmol). The reaction mixture was stirred at 0°C for 1 h. EDC (0.043 g, 0.215 mmol) and triethylamine (0.060 mL, 0.430 mmol) were added and the mixture was stirred at room temperature for 16 h. The reaction was poured into water (10 mL) and stirred. The resulting solid was filtered and dried in a vacuum oven at 50 °C overnight to afford (E)-1-(4-(4-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide (0.043 g, 0.056 mmol, 79% yield). LCMS ( m/z ): 740.5 [M + H] ⁺ .

Step 5: (E)-1-((E)-4-((E)-4-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To (E)-1-(4-(4-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide (0.040 g, 0.054 mmol) in DMF (0.54 mL) was added cesium carbonate (0.053 g, 0.162 mmol) and iodomethane (7.4 µL, 0.12 mmol). The mixture was stirred at room temperature for 16 h. The mixture was partitioned between ethyl acetate (10 mL) and brine (5 mL). The aqueous layer was extracted with EtOAc (10 mL). The combined EtOAc layers were dried over magnesium sulfate, filtered and concentrated. The residue was purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 30-85% gradient of MeCN/water with 0.075% _NH4OH , 10 mM ammonium bicarbonate, pH 10) to afford (E)-1-((E)-4-((E)-4-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (0.013 g, 0.016 mmol, 30% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.11 (d, J =1.27 Hz, 1 H), 8.04 (br. s., 1 H), 7.80 (dd, J =1.39, 8.49 Hz, 1 H), 7.49-7.53 (m, 1 H), 7.46 (br. s., 1 H) ), 7.28 (t, J =8.49 Hz, 2 H), 7.11-7.18 (m, 1 H), 6.45 (s, 1 H), 6.42 (s, 1 H), 4.85 (br. s., 4 H), 4.51 (dq, J =3.68, 6.97 Hz, 4 H), 3.78 (s, 3 H) , 3.59 (s, 3 H), 2.128 (s, 3 H), 2.125 (s, 3 H), 1.59 (s, 6 H), 1.25-1.35 (m, 6 H). LCMS ( m/z ): 768.4 [M + H] ⁺ .

Examples 32 and 33

Example 32: (E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropyloxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (earlier eluting enantiomer)

Example 33: (E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropyloxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (later eluting enantiomer)

Step 1: (E)-1-(4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (10.4 g, 21.47 mmol) in 1-butanol (150 mL) at room temperature was added DIEA (7.50 mL, 42.9 mmol). The reaction mixture was then stirred at room temperature for 2 h. 3-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-chloro-5-nitrobenzamide (10.9 g, 28.0 mmol) was added, followed by sodium bicarbonate (5.41 g, 64.4 mmol). The reaction mixture was then stirred at 100 °C for 4 days. The reaction mixture was cooled to room temperature and concentrated. The resulting orange sludge was suspended in acetonitrile and then filtered. The solid was washed with acetonitrile and water. Then, the solid was dried to obtain (E)-1-(4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (10.94 g, 14.32 mmol, 67% yield) as an orange-red solid. ¹ H NMR (400MHz, DMSO- d ₆ ) δ ppm 13.27 - 12.18 (m, 1 H), 8.14 (d, J =1.8 Hz, 1 H), 7.97 (br. s., 2 H), 7.78 - 7.62 (m, 2 H), 7.49 (d, J =1.8 Hz, 1 H), 7 .38 - 7.24 (m, 3 H), 6.60 (s, 1 H), 5.92 - 5.63 (m, 2 H), 4.90 (d, J =5.3 Hz, 2 H), 4.58 (q, J =7.1 Hz, 2 H), 4.13 (t, J =5.4 Hz, 2 H), 4.00 (t, J =6.0 Hz, 2H), 3.87 (s, 3 H), 3.64 (t, J =6.1 Hz, 2 H), 2.16 (s, 3H), 1.82 (quint, J =6.0 Hz, 2 H), 1.33 (t, J =7.0 Hz, 3 H), 0.79 (s, 9 H), -0.05 (s, 6 H). LCMS ( m/z ): 764.7 [M + H] ⁺ .

Step 2: (E)-1-(4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

To a solution of (E)-1-(4-((2-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoyl-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (10.1 g, 13.22 mmol) in methanol (200 mL) stirred at 60 °C was added a solution of sodium dithionite (25.0 g, 121 mmol) in water (200 mL). The reaction mixture was then stirred at the same temperature for 1 h. The reaction mixture was then cooled to room temperature and quenched with 500 mL of water. The resulting mixture was filtered. The collected solid was washed with water (500 mL x 3) and then rinsed with diethyl ether (300 mL). The solid was then dried in a vacuum oven to afford partially pure (E)-1-(4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (7.36 g, ~10 mmol, ~76% yield) as a light brown solid. The approximate purity of the title compound by LCMS was 63% (UV, m/z = 734.6 [M + H] ⁺ ) and 20% of the silyl-deprotected byproduct (UV, m/z = 620.5 [M + H] ⁺ ). The mixture was used in the next reaction without further purification.

Step 3: (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To a solution of (E)-1-(4-((2-amino-6-(3-((tert-butyldimethylsilyl)oxy)propoxy)-4-carbamoylphenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (7.36 g, 10.03 mmol, ~76% purity) in DMF (60 mL) was added a 1 M solution of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (15.1 mL, 15.10 mmol) in 1,4-dioxane dropwise via an addition funnel at room temperature. The reaction mixture was then stirred at room temperature for 1 h. Then, EDC (3.84 g, 20.06 mmol) and TEA (5.6 mL, 40.2 mmol) were added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 48 h. The reaction mixture was diluted with EtOAc and washed with water. Solids were removed by filtration and identified as alcohol derivatives of silyl-deprotection. Then, the organic layer was washed for the second time. The water layer merged was extracted with EtOAc (1x). The organic layer merged was washed with salt water, dried over magnesium sulfate and concentrated. The crude product and solid separated earlier in the process were suspended in EtOH and filtered. The solid was washed with EtOH, then dried to obtain a pale solid (6.0 g). A second batch of solids (824 mg) was also obtained. The combined mixture (6.824 g, ~7.6 mmol, ~76%) was characterized by LCMS and NMR as a roughly 3:1 mixture of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (LCMS ( m/z ): 895.8 [M + H] ⁺ ) and the alcohol derivative (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-5-carboxamide (LCMS ( m/ z ): 781.7 [M+H] ⁺ ). The mixture was used in the next reaction without further purification.

Step 4: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2-hydrochloride

To a solution of (E)-7-(3-((tert-butyldimethylsilyl)oxy)propoxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (6.0 g, 6.70 mmol, as a 3:1 mixture of silyl ethers/alcohols) in THF (50 mL) was added 4M HCl solution in dioxane (8.4 mL, 33.6 mmol) at room temperature. The reaction mixture was then stirred at room temperature for 5 h. Additional HCl solution (4.2 mL, 16.80 mmol) was added and the reaction mixture was stirred overnight. Add additional HCl solution (4.2 mL, 16.80 mmol) and stir the reaction mixture for 24 h. Add additional HCl solution (8.4 mL, 33.6 mmol). Stir the mixture at room temperature for 1 h and then at 40 °C for 5 h. Add additional HCl solution (8.4 mL, 33.6 mmol) and stir the reaction mixture at 40 °C over the weekend. Cool the reaction mixture to room temperature and then filter. The solid was washed with THF and dried to afford (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (5.6 g, 6.56 mmol, 98% yield) as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ) δ ppm 7.98 (br. s., 2 H), 7.65 (dd, J =1.1, 3.6 Hz, 2 H), 7.33 (s, 4 H), 6.53 (d, J =1.5 Hz, 2 H), 5.86 - 5.81 (m, 2H), 4.9 3 (dd, J =3.8, 6.8 Hz, 4 H), 4.57 - 4.47 (m, 4 H), 4.07 (t, J =6.4 Hz, 2H), 3.75 (s, 3 H), 3.49 - 3.42 (m, 2 H), 2.11 (two s, 6 H), 1.80 - 1.64 (m, 2H), 1.27 (two t, J =7.2 Hz, 6 H). LCMS ( m/z ): 781.7 [M + H] ⁺ .

Step 5: (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, 2 hydrochloride (5.45 g, 6.38 mmol) and cesium carbonate (10.40 g, 31.9 mmol) in DMF (35 mL) was added iodomethane (0.918 mL, 14.68 mmol) by using an ice bath at 0°C. The ice bath was removed and the mixture was stirred at room temperature for 16 h. The mixture was partitioned between water and 3:1 chloroform/ethanol. The layers were separated. The aqueous layer was extracted with 3:1 chloroform/ethanol (6X). The organic layer was washed with water. The aqueous wash solution was extracted again with 3:1 chloroform/ethanol. The combined organic layers were dried over sodium sulfate, filtered, and concentrated to provide the title compound (6 g). The approximate purity of the title compound measured by LCMS was 76% (UV210-350nm, m/z =809.3 [M + H] ⁺ ). This compound was used in the next reaction without further purification.

Step 6: (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropyloxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (5.2 g, 6.43 mmol) in tert-butanol (52 mL) and water (13 mLl) was added NMO (1.130 g, 9.64 mmol). After stirring at room temperature for 5 min, 2.5% osmium tetroxide (4.04 mL, 0.321 mmol) in the tert-butyl alcohol is added and the mixture is stirred for 18 h. After concentration, the residue is suspended in water and filtered. The collected solid is rinsed with water and dried in a vacuum to provide (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo [d] imidazoles-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazoles-5-carbonyl) imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo [d] imidazoles-5-carboxamide (4.0 g, 73.8% yield) as a mixture of stereoisomers. LCMS (~90% purity by UV 210-350 nm; m/z ): 843.2 [M + H] ⁺ .

Step 7: Earlier eluting enantiomer Example 33 (E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropyloxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide and later eluting enantiomer Example 34 Purification of (E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

For registration and screening, 1500 mg portions of the crude product were separated into the individual enantiomers by a sequence with two preparative HPLC purification steps.

First purification step

The purpose of this step is to separate the racemic mixture of the desired enantiomer from several minor byproducts (eluting near the co-eluting pair of enantiomers). Use the following method:

。

.

Results: The purest fractions were combined and concentrated to a volume of 10 mL. The resulting precipitate was filtered and dried at 35 °C to provide a racemic mixture of enantiomers (600 mg, 0.71 mmol). The C18 HPLC method (Method 2) was also used to demonstrate the purification effect (purity 98.95%). LCMS ( m/z ): 843.3 [M+H] ⁺ . Similar treatment of fractions with slightly lower purity (i.e., front fraction and tail fraction) provided an additional amount of a racemic mixture of enantiomers (310 mg).

Second purification step: The purpose of this step is to separate and isolate the individual enantiomers. The following method was used:

。

.

Results: The pure fractions of each enantiomer were concentrated and dried under high vacuum at 40°C to afford the following solid. The assignment of absolute stereochemistry can be obtained by subsequent re-synthesis from chiral building blocks.

Example 32 (first eluting isomer):

(E)-1-((2R,3R)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (247 mg, 0.293 mmol) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δppm 1.26 (t, J =7.1 Hz, 3H) 1.27 (t, J =7.1 Hz, 3H) 1.83 (quint, J =6.15 Hz, 2H) 2.10 (s, 3 H) 2.11 (s, 3 H) 3.50 (s, 3 H) 3.50 - 3.55 (m, 5 H) 3.80 (s, 3H) 3.81 - 3.91 (m, 2 H) 4.18 (t, J =6.46 Hz, 2 H) 4.24 - 4.33(m, 2 H) 4.44 -4.53 (m, 5 H) 4.53 - 4.60 (m, 2 H) 4.97 (d, J =6.84 Hz, 1 H) 5.06 (d, J =6.34 Hz, 1 H) 6.41 (s, 1 H) 6.44 (s, 1 H) 7.41 - 7.50 (m, 4 H) 7.71 (dd, J =2.66,1.14 Hz, 2 H) 8.06 (br. s., 2 H). LCMS (100% purity by UV210-350 nm; m/z ): 843.3 [M + H] ⁺ . Enantiomeric excess > 98 % ee by chiral analytical HPLC. αD ²⁰ - 24 ( c 0.1, MeOH).

Example 33 (second eluting isomer):

(E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (227 mg, 0.269 mmol) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δppm 1.26 (t, J =7.1 Hz, 3 H) 1.27 (t, J =7.1 Hz, 3 H) 1.83 (quint, J =6.21 Hz, 2 H) 2.10 (s, 3 H) 2.11 (s, 3 H) 3.50 (s, 3 H) 3.50 - 3.56 (m, 5 H) 3.80 (s, 3 H) 3.81 - 3.92 (m, 2 H) 4.18 (t, J =6.34 Hz, 2 H) 4.24 - 4.33 (m, 2 H) 4.44- 4.52 (m, 5 H) 4.53 - 4.61 (m, 2 H) 4.97 (d, J =6.84 Hz, 1 H) 5.06 (d, J =6.34 Hz, 1 H) 6.41 (s, 1 H) 6.44 (s, 1 H) 7.41 - 7.50 (m, 4 H) 7.71 (dd, J =2.53,1.01 Hz, 2 H) 8.06 (br. s., 2 H). LCMS (100% purity by UV210-350 nm; m/z ): 843.3 [M + H] ⁺ . Enantiomeric excess 98 % ee by chiral analytical HPLC. αD ²⁰ + 23 ( c 0.1, MeOH).

Embodiment 34

(E)-1-(4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: tert-Butyl (E)-(4-((2-bromo-6-nitrophenyl)amino)but-2-en-1-yl)carbamate

To a solution of (E)-(4-aminobutan-2-ene-1-yl)t-butylcarbamate (2.2 g, 11.81 mmol) and DIEA (4.37 mL, 25.00 mmol) in isopropanol (30 mL) was added 1-bromo-2-fluoro-3-nitrobenzene (2.5 g, 11.36 mmol) at 25 ° C. Then, the reaction mixture was stirred at 25 ° C for 4 days. The reaction mixture was concentrated. The resulting material was distributed between water and EtOAc. The aqueous layer was separated and extracted with EtOAc (1x). Then, the combined organic layer was washed with brine, dried over magnesium sulfate, and concentrated to obtain (E)-(4-((2-bromo-6-nitrophenyl)amino)butan-2-ene-1-yl)t-butylcarbamate (4.6 g, 12 mmol, 100% yield) as a yellow solid. The isolated material was used without any further purification. LCMS ( m/z ): 332.0 ([M+H] ⁺ -tert-butyl).

Step 2: tert-Butyl (E)-(4-((2-amino-6-bromophenyl)amino)but-2-en-1-yl)carbamate

To a mixture of tert-butyl (E)-(4-((2-bromo-6-nitrophenyl)amino)but-2-en-1-yl)carbamate (4.4 g, 11.39 mmol) and ammonium chloride (6.09 g, 114 mmol) in methanol (50 mL) was added zinc (7.45 g, 114 mmol). Then, the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was filtered and concentrated. The separated residue was distributed between EtOAc and water. The aqueous layer was separated and the organic layer was washed with water for a second time. The combined aqueous layer was extracted with EtOAc (1x). Then, the combined organic layer was washed with brine, dried over magnesium sulfate, filtered and concentrated to obtain the title compound (4.0 g, ~11.23 mmol) as a light brown oil. LCMS (77% purity measured by UV210-350nm; m/z ): 356.1 [M + H] ⁺ . The product was used without any further purification.

Step 3: tert-Butyl (E)-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

To a solution of tert-butyl (E)-(4-((2-amino-6-bromophenyl)amino)but-2-en-1-yl)carbamate (4.0 g, 11.23 mmol) in DMF (40 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate solution (1 M in 1,4-dioxane, 12.4 mL, 12.40 mmol) at room temperature. The reaction mixture was then stirred for 45 min. EDC (3.23 g, 16.84 mmol) and TEA (4.7 mL, 33.7 mmol) were then added to the reaction mixture at room temperature. After stirring for 2 h, the mixture was diluted with EtOAc and washed with water (2x). The combined aqueous layer was extracted again with EtOAc (1x). The combined organic layer was washed with brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by normal phase chromatography (ISCO CombiFlash, 120 g Gold column, DCM/MeOH) to afford the title compound as an off-white solid after solvent evaporation (4.5 g, 8.70 mmol, 77% yield). LCMS ( m/z ): 517.2 [M + H] ⁺ .

Step 4: (E)-N-(1-(4-aminobut-2-en-1-yl)-7-bromo-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide, hydrochloride

To a solution of tert-butyl (E)-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (1.1 g, 2.126 mmol) in methanol (10 mL) was added HCl (4 M in dioxane, 5.00 mL, 20 mmol) at room temperature. The reaction mixture was then stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and suspended in diethyl ether. The resulting solid was filtered, washed with diethyl ether, and then dried to afford (E)-N-(1-(4-aminobut-2-en-1-yl)-7-bromo-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide, hydrochloride (1.06 g) as a white solid. The isolated material was used without any further purification. LCMS ( m/z ): 417.1 [M+H] ⁺ .

Step 5: (E)-N-(7-Bromo-1-(4-((4-carbamoyl-2-nitrophenyl)amino)but-2-en-1-yl)-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide

To a suspension of (E)-N-(1-(4-aminobut-2-en-1-yl)-7-bromo-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide, hydrochloride (0.965 g, 2.126 mmol) and 4-fluoro-3-nitrobenzamide (0.431 g, 2.339 mmol) in isopropanol (10 mL) was added DIEA (0.780 mL, 4.46 mmol) at room temperature. The reaction mixture was stirred at 70°C overnight. Then, the reaction mixture was cooled to room temperature and the solid was collected on a filter. The solid was washed with isopropanol and dried to afford (E)-N-(7-bromo-1-(4-((4-carbamoyl-2-nitrophenyl)amino)but-2-en-1-yl)-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide (1.2 g, 2.06 mmol, 97% yield) as a yellow solid. The isolated material was used without any further purification. LCMS ( m/z ): 581.1 [M+H] ⁺ .

Step 6: (E)-N-(1-(4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)-7-bromo-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide

To a solution of (E)-N-(7-bromo-1-(4-((4-carbamoyl-2-nitrophenyl)amino)but-2-en-1-yl)-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide (1.2 g, 2.064 mmol) and ammonium chloride (1.1 g, 20.56 mmol) in methanol (15 mL) was added zinc (1.3 g, 19.88 mmol). The reaction mixture was then stirred at room temperature for 8 h. Another 10 eq of ammonium chloride (1.1 g, 20.56 mmol) and zinc (1.3 g, 19.88 mmol) were added and the mixture was stirred at room temperature overnight. As the reduction was still not complete, acetic acid (1.5 mL) was added and then stirred at room temperature for 30 min.

The reaction mixture is filtered and the filtrate is concentrated. The separated material is distributed between EtOAc and water. The solid that appears is collected on the filter and washed with EtOAc and water. Then, the filtrate is placed in a separatory funnel. The aqueous layer is extracted with EtOAc (1x). Then, the organic layer combined is washed with salt water, dried (MgSO ) and concentrated. The above separated residue and solid are dissolved in MeOH and evaporated onto diatomaceous earth. Silica gel chromatography (dry loading, 40 g column, 0-10% gradient of MeOH/DCM) afforded an off-white solid (911 mg, ~1.6 mmol) as a ~3:1 mixture of (E)-N-(1-(4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)-7-bromo-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide (LCMS ( m/z ): 551.2 [M+H] ⁺ ) and debrominated byproducts. The isolated material was used directly in the next reaction.

Step 7: (E)-1-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To a solution of (E)-N-(1-(4-((2-amino-4-carbamoylphenyl)amino)but-2-en-1-yl)-7-bromo-1H-benzo[d]imidazol-2-yl)-1-ethyl-3-methyl-1H-pyrazole-5-carboxamide (911 mg, ~1.6 mmol, with ~25% deficient bromine atoms) in DMF (10 mL) was added a 1 M solution of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (1.82 mL, 1.82 mmol) in 1,4 dioxane at room temperature. The reaction mixture was then stirred at room temperature for 1.5 h. EDC (633 mg, 3.30 mmol) and TEA (0.921 mL, 6.61 mmol) were then added at room temperature. After stirring at room temperature overnight, the reaction mixture was diluted with EtOAc and water. The suspended solid was filtered and washed with EtOAc, water, EtOAc, and then diethyl ether. The off-white solid was dried to provide a 85:15 mixture of (E)-1-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (883 mg, ~1.2 mmol, 75% yield, LCMS ( m/z ): 712.2 [M+H] ⁺ ) and the corresponding des-bromide analog. The material was used without further purification.

Step 8: (E)-1-((E)-4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a mixture of (E)-1-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (816 mg, 1.145 mmol, ~15% debrominated impurity) and cesium carbonate (1.5 g, 4.60 mmol) in DMF (10 mL) was added iodomethane (0.143 mL, 2.29 mmol) at room temperature. The reaction was then stirred at room temperature for 4 h. Additional iodomethane (0.143 mL, 2.290 mmol) was added and the reaction mixture was stirred at room temperature for 45 h. Water and EtOAc were added to produce a solid (345 mg, unreacted starting material). The filtrate is separated and aqueous phase is extracted with EtOAc.The organic layer merged is washed with salt water, dried over magnesium sulfate, and concentrated.The (E)-1-((E)-4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino)-3-methyl-2,3-dihydro-1H-benzo [d] imidazoles-1-yl) but-2-ene-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazoles-5-carbonyl) imino)-3-methyl-2,3-dihydro-1H-benzo [d] imidazoles-5-carboxamide (107 mg, 0.144 mmol, 12.6% yield) as white solid is provided by the purification (30 mm x 50 mm GeminiC18, gradient ACN/ water, containing 0.1% TFA modifier) of preparative HPLC. ¹ H NMR (400MHz, DMSO- d ₆ ) δppm 8.09 (d, J =1.3 Hz, 1 H), 8.03 (br. s., 1 H), 7.81 (dd, J =1.5, 8.3 Hz, 1H), 7.60 (dd, J =0.8, 8.0 Hz, 1 H), 7.50 (d, J = 8.3 Hz, 1 H), 7.45 (dd, J =0.8,8.0 Hz, 2 H), 7.23 (t, J =8.2 Hz, 1 H), 6.45 (s, 1 H), 6.39 (s, 1 H), 5.95 (td, J =4.8, 15.7 Hz, 1 H), 5.56 (td, J =5. 8, 15.7 Hz, 1 H), 5.01 (d, J =3.8 Hz,2 H), 4.78 (d, J =5.5 Hz, 2 H), 4.49 - 4.38 (m, 4 H), 3.97 (s, 2 H), 3.51 (s, 3 H), 2.13 (s, 3 H), 2.11 (s, 3 H), 1.1 5-1.28 (m, 6 H). LCMS ( m/z ): 740.3 [M +H] ⁺ .

Step 9: (E)-1-(4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-((E)-4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (54 mg, 0.073 mmol) in tert-butyl alcohol (0.8 mL) and water (0.2 mL) was added NMO (26 mg, 0.22 mmol). After stirring at room temperature for 5 min, 2.5% osmium tetroxide in tert-butyl alcohol (0.183 mL, 0.015 mmol) was added and stirring was continued at room temperature for 2 h. The reaction mixture was filtered and the filtrate was directly purified by reverse phase HPLC (30 mm x 50 mm Gemini C18, gradient of ACN/water with 0.1% ammonium hydroxide modifier). (E)-1-(4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (37 mg, 0.48 mmol, 65% yield) was obtained as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ) δ ppm 8.09 -8.00 (m, 2 H), 7.85 (dd, J =1.5, 8.4 Hz, 1 H), 7.61 - 7.56 (m, 2 H), 7.48 -7.39 (m, 2 H), 7.23 (t, J =8.0 Hz, 1 H ), 6.47 (d, J =7.1 Hz, 2 H), 5.36 (d, J =6.6 Hz, 1 H), 5.11 (d, J =6.6 Hz, 1 H), 4.71 (dd, J =9.1, 14.4 Hz, 1 H), 4.50 (quint, J =6.7 Hz, 4 H), 4.38 (dd, J =3.8, 14.4 Hz, 1 H), 4.30 - 4.15 (m, 2H), 4.04 - 3.89 (m, 2 H), 3.57 (s, 3 H), 3.51 (s, 3 H), 2.11 (s, 3 H), 2.15 (s, 3 H), 1.34 - 1.21 (m, 6 H). LCMS ( m/z ): 774.3 [M + H] ⁺ ).

Embodiment 35

(2E,2'E)-1,1'-((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide)

Step 1: 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide)

To a mixture of 4-fluoro-3-nitrobenzamide (0.985 g, 5.35 mmol) in 1-butanol (10 mL) was added (2S,3S)-2,3-diethoxybutane-1,4-diamine (0.46 g, 2.61 mmol) and DIEA (1.82 mL, 10.4 mmol). The mixture was stirred at 110 °C for 2 h. When cooled to room temperature, the solid was collected on a filter, washed with a mixture of diethyl ether and 2-propanol (1:1) and dried to provide 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide) (0.63 g, 1.25 mmol, 48% yield) as an orange solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 1.13 (t, J =7.0 Hz, 6 H), 3.45 - 3.74 (m, 8H), 3.79 - 3.89 (m, 2 H), 7.17 (d, J =9.1 Hz, 2 H), 7.31 (br. s., 2 H), 8 .02(dd, J =8.9, 2.0 Hz, 4 H), 8.56 (t, J =5.2 Hz, 2 H), 8.66 (d, J =2.0 Hz, 2 H). LCMS ( m/z ): 505.1 [M + H] ⁺ .

Step 2: 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide)

To a 100 mL round bottom flask was added 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-nitrobenzamide) (0.63 g, 1.25 mmol) and methanol (20 mL). To this mixture was added 10 mL of saturated aqueous ammonium chloride solution. To this mixture was added zinc (0.812 g, 12.5 mmol) and the heterogeneous mixture was stirred at room temperature for 15 min. The mixture was passed through a filter and rinsed with MeOH and then concentrated. Silica gel chromatography (24 g column, 6-20% gradient of MeOH/DCM with 1% ammonium hydroxide as modifier) provided 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide) (0.446 g, 1.00 mmol, 80% yield) as a light yellow solid. LCMS ( m/z ): 445.4 [M+H] ⁺ .

Step 3: 1,1'-((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide)

To a solution of 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-aminobenzamide) (0.446 g, 1.00 mmol) in DMF (20 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~0.4 M in dioxane, 5.02 mL, 2.00 mmol). The mixture was stirred for 15 min. EDC (0.481 g, 2.51 mmol) and TEA (0.699 mL, 5.02 mmol) were added and the reaction mixture was stirred at room temperature for 18 h. The mixture was poured into 3:1 water:saturated aqueous ammonium chloride solution (100 mL). A fine solid was formed immediately and stirring was continued for another 10 min. The resulting solid was filtered, washed with water, and dried to provide 1,1′-((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (0.539 mg, 0.701 mmol, 70% yield) as a white solid. LCMS ( m/z ): 767.5 [M+H] ⁺ .

Step 4: (2E,2'E)-1,1'-((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide)

To a solution of 1,1'-((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide) (0.09 g, 0.117 mmol) in DMF (5 mL) was added cesium carbonate (0.103 g, 0.317 mmol) and iodomethane (0.018 mL, 0.282 mmol). The reaction mixture was stirred at room temperature for 18 h. More cesium carbonate (0.019 g, 0.059 mmol) and iodomethane (0.015 mL, 0.235 mmol) were added. The mixture was stirred at 50 ^°C for 1 h. The reaction was diluted with water and extracted with EtOAc (3 x 50 mL). The organic phase was washed with brine (10 mL), dried over magnesium sulfate and concentrated. Use mass-guided HPLC (XSelect CSHPrep C18, 5 um, 15-55% gradient ACN/ water, containing 0.1% TFA as modifier) purified product. Combine each fraction and remove ACN. Use saturated ammonium bicarbonate solution to alkalize aqueous phase. Filter gained solid and dry on freeze dryer to provide (2E, 2'E)-1,1'-((2S, 3S)-2,3-diethoxybutane-1,4-diyl)bis-(2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino)-3-methyl-2,3-dihydro-1H-benzo [d] imidazole-5-carboxamide) (13 mg, 0.016mmol, 14% yield) as white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.01 - 8.15 (m, 4 H) 7.80 -7.93 (m, 2 H) 7.61 (d, J =8.36 Hz, 2 H) 7.46 (br. s., 2 H) 6.51 (s, 2 H) 4.46- 4.62 (m, 6 H) 4.38 (dd, J =14.45, 8.87 Hz, 2 H) 3.78 - 3.93 (m, 2 H) 3.60 (s, 6 H) 3.25 - 3.33 (m, 2 H) 3.02 (dd, J =9.38, 7.10 Hz, 2 H) 2.12 (s, 6 H)1.31 (t, J =7.10 Hz, 6 H) 0.59 (t, J =6.97 Hz, 6 H). LCMS ( m/z ): 795.3 [M + H] ⁺ .

Embodiment 36

(E)-1-(((4R,5R)-5-(((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: 4-((((4R,5R)-5-(aminomethyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-3-(3-(benzyloxy)propoxy)-5-nitrobenzamide

To a solution of ((4R,5R)-2,2-dimethyl-1,3-dioxolane-4,5-diyl)dimethylamine (1 g, 6.24 mmol) and 3-(3-(benzyloxy)propoxy)-4-chloro-5-nitrobenzamide (2.049 g, 5.62 mmol) in 1-butanol (20 mL) was added DIEA (3.27 mL, 18.72 mmol). The reaction mixture was stirred at 120 °C for 16 h. The mixture was concentrated under vacuum to provide the crude product. The crude product was purified by silica gel chromatography (elution gradient: 0 to 30% MeOH in DCM). Pure fractions were evaporated to dryness to afford 4-((((4R,5R)-5-(aminomethyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-3-(3-(benzyloxy)propoxy)-5-nitrobenzamide (1.5 g, 2.76 mmol, 44.3% yield) as a red gum. LCMS ( m/z ): 489 [M+H] ⁺ .

Step 2: 3-(3-(Benzyloxy)propoxy)-4-((((4R,5R)-5-(((4-carbamoyl-2-nitrophenyl)amino)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-5-nitrobenzamide

To a solution of 4-((((4R,5R)-5-(aminomethyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-3-(3-(benzyloxy)propoxy)-5-nitrobenzamide (1.4 g, 2.87 mmol) and 4-fluoro-3-nitrobenzamide (0.580 g, 3.15 mmol) in DMSO (15 mL) was added K2CO3 (0.792 g, 5.73 mmol). The reaction mixture was stirred at 25 °C for 16 h. The mixture was poured into water (100 mL). The precipitate was collected by filtration, washed with water and dried under vacuum to afford 3-(3-(benzyloxy)propoxy)-4-((((4R,5R)-5-(((4-carbamoyl-2-nitrophenyl)amino)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-5-nitrobenzamide (1.4 g, 1.931 mmol, 67.4% yield) as an orange solid. LCMS (m/z): 653 [M +H] ⁺ .

Step 3: 3-amino-4-((((4R,5R)-5-(((2-amino-4-carbamoylphenyl)amino)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-5-(3-(benzyloxy)propoxy)benzamide

To a solution of 3-(3-(benzyloxy)propoxy)-4-((((4R,5R)-5-(((4-carbamoyl-2-nitrophenyl)amino)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-5-nitrobenzamide (1.35 g, 2.068 mmol) in acetic acid (20 mL) was added zinc (1.352 g, 20.68 mmol). The reaction mixture was stirred at 25 °C for 3 h. The mixture was diluted with DCM (50 mL) and filtered. The filtrate was concentrated under reduced pressure to afford crude 3-amino-4-((((4R,5R)-5-(((2-amino-4-carbamoylphenyl)amino)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-5-(3-(benzyloxy)propoxy)benzamide (1.3 g, 1.755 mmol, 85% yield) as a grey solid. LCMS (m/z): 593 [M + H] ⁺ .

Step 4: 2-amino-1-(((4R,5R)-5-((2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-7-(3-(benzyloxy)propoxy)-1H-benzo[d]imidazole-5-carboxamide

To a solution of 3-amino-4-((((4R,5R)-5-(((2-amino-4-carbamoylphenyl)amino)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)amino)-5-(3-(benzyloxy)propoxy)benzamide (1.35 g, 2.278 mmol) in methanol (20 mL) was added cyanogen bromide (0.724 g, 6.83 mmol). The reaction mixture was stirred at 25 °C for 16 h. The mixture was diluted with diethyl ether (30 mL). The mixture was filtered and washed with diethyl ether. The filtrate was concentrated under reduced pressure to afford 2-amino-1-(((4R,5R)-5-((2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-7-(3-(benzyloxy)propoxy)-1H-benzo[d]imidazole-5-carboxamide (800 mg, 1.12 mmol, 49.2% yield) as a grey solid. LCMS (m/z): 643 [M + H] ⁺ .

Step 5: 7-(3-(Benzyloxy)propoxy)-1-(((4R,5R)-5-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide

To a mixture of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (336 mg, 2.178 mmol), 2-amino-1-(((4R,5R)-5-((2-amino-5-carbamoyl-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-7-(3-(benzyloxy)propoxy)-1H-benzo[d]imidazole-5-carboxamide (700 mg, 1.089 mmol) and DIPEA (0.951 mL, 5.45 mmol) in DMF (10 mL) was added HATU (1035 mg, 2.72 mmol). The reaction mixture was stirred at 60° C. for 16 h. The mixture was poured into water. The precipitate was collected by filtration, washed with water and diethyl ether, and then dried under vacuum to afford 7-(3-(benzyloxy)propoxy)-1-(((4R,5R)-5-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (800 mg, 0.743 mmol, 68.2% yield) as a brown solid. LCMS (~85% purity by UV, m/z ): 915 [M + H] ⁺ .

Step 6: 1-(((4R,5R)-5-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazol-5-carboxamide

To a solution of 7-(3-(benzyloxy)propoxy)-1-(((4R,5R)-5-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (650 mg, 0.710 mmol) in methanol (20 mL) was added Pd—C (756 mg, 7.10 mmol). The reaction was hydrogenated at 60° C. using H-cube (4 atm) for 72 h. The mixture was diluted with DMF (20 mL). The mixture was filtered and the filtrate was concentrated under reduced pressure to provide the crude product. The crude product was purified by preparative HPLC (Gemini-C18 column, 5µ silica, 21 x 150 mm; 30-40% gradient of ACN/water with 0.1% TFA modifier) to afford 1-(((4R,5R)-5-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazole-5-carboxamide (30 mg, 0.035 mmol, 4.9% yield) as an off-white solid. ¹ H NMR (400 MHz, MeOH- d4 ) δ 7.58 (s, 1 H), 7.48 (d, J = 8.4 Hz, 1 H), 7.33 (s, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 6.89 (s, 1 H), 6.58 (s, 2 H), 5.11- 4.99 (m, 1 H), 4.93 (s, 1 H), 4.61 (dqd, J = 26.0, 13.2, 6.9 Hz, 5 H), 4.46- 4.36 (m, 1 H), 4.31 (dd, J = 13.4, 3.2 Hz, 1 H), 4.19 (dd, J = 14.0, 3.4Hz, 1 H), 4.03 (dd, J = 15.1, 6.4 Hz, 1 H), 3.88 (dd, J = 14.9, 6.4 Hz, 1 H), 3.83 - 3.72 (m, 1 H), 2.23 (s, 3 H), 2.18 (s, 3 H), 2.05 (dd, J = 11.5, 5.9 Hz, 2 H), 1.64 (d, J = 10.3 Hz, 6 H), 1.50 - 1.28 (m, 6 H). LCMS ( m/z ): 825 [M+ H] ⁺ .

Step 7: (E)-1-(((4R,5R)-5-(((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To an ice-bath cooled mixture of 1-(((4R,5R)-5-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)methyl)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-hydroxypropoxy)-1H-benzo[d]imidazole-5-carboxamide (21 mg, 0.025 mmol) and cesium carbonate (24.88 mg, 0.076 mmol) in DMF (0.5 mL) was added iodomethane (4 µL, 0.064 mmol). The ice bath was removed and the mixture was stirred at room temperature for 16 h. After filtration, the filtrate was directly purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 30-85% gradient of MeCN/10 mM ammonium bicarbonate, adjusted to pH 10 with ammonia). Concentration of pure fractions afforded the title compound (12 mg, 0.014 mmol, 55% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 1.28 - 1.35 (m,12 H) 1.89 - 1.94 (m, 2 H) 2.08 (s, 3 H) 2.13 (s, 3 H) 3.28 (s, 3 H) 3.30 (s,3 H) 3.53 - 3.57 (m, 2 H) 4.01 - 4.07 (m, 1 H) 4.11 - 4.16 (m, 1 H) 4.27 -4.31 (m, 2 H) 4.37 (dd, J =12.67, 4.31 Hz, 1 H) 4.49 - 4.56 (m, 6 H) 4.60 -4.66 (m, 2 H) 6.59 (d, J =7.10 Hz, 2 H) 7.27 (s, 1 H) 7.42 - 7.50 (m, 4 H)7.72 (dd, J =8.49, 1.39 Hz, 1 H) 7.79 (s, 1 H) 8.05 (d, J =16.48 Hz, 2 H). LCMS ( m/z ): 853.4 [M + H] ⁺ .

Embodiment 37

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-isopropyl-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-bromo-5-nitrobenzamide

To a solution of 3-bromo-4-fluoro-5-nitrobenzamide (1.4 g, 5.32 mmol) and 2,2,3,3-tetrafluorobutane-1,4-diamine, 2 hydrochloride (1.3 g, 5.58 mmol) in ethanol (30 mL) was added DIEA (3.53 mL, 20.23 mmol) at room temperature. Then, the reaction mixture was warmed to 70 ° C and stirred for 4 h. The reaction mixture was cooled to room temperature and then concentrated. The resulting material was distributed between water and EtOAc. The aqueous layer was separated and extracted with EtOAc (1x). Then, the combined organic layer was washed with brine, dried with magnesium sulfate and concentrated to obtain 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-bromo-5-nitrobenzamide (2.15 g, 5.3 mmol, 100% yield) as a yellow solid. The solid was used without further purification. LCMS ( m/z ): 403.0 [M + H] ⁺ .

Step 2: 3-Bromo-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-nitrobenzamide

To a suspension of 4-fluoro-3-nitrobenzamide (1.5 g, 8.15 mmol) and 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-bromo-5-nitrobenzamide (2.15 g, 5.3 mmol) in ethanol (25 mL) was added DIEA (2.8 mL, 16.03 mmol) at room temperature. The reaction mixture was then warmed to 80 °C and stirred for 48 h. The reaction mixture was then cooled to room temperature and filtered. The solid was washed with EtOH and then dried to obtain 3-bromo-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-nitrobenzamide (2.6 g, 4.58 mmol, 86% yield) as a yellow solid. LCMS ( m/z ): 567.0 [M + H] ⁺ .

Step 3: 4-((4-((4-Carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-3-nitro-5-(prop-1-en-2-yl)benzamide

To a 40-mL scintillation vial containing 3-bromo-4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-nitrobenzamide (300 mg, 0.529 mmol), trifluoro(prop-1-en-2-yl)-14-borane, potassium salt (196 mg, 1.322 mmol) and K ₃ PO ₄ (393 mg, 1.851 mmol) in DMF (2.5 mL) and water (0.25 mL) was added PdCl ₂ (dppf)-CH ₂ Cl adduct (44 mg, 0.054 mmol) at room temperature. The reaction vessel was then evacuated and backfilled with nitrogen. The reaction mixture was then warmed to 80 °C and stirred overnight. When cooled to room temperature, the mixture was diluted with EtOAc and water. The two-phase mixture is filtered through a diatomaceous earth pad. Then, the aqueous layer is separated and the organic layer is washed with water (twice again). The aqueous layer merged is then extracted with EtOAc (1x). The organic layer merged is washed with saturated brine, dried over magnesium sulfate and concentrated. The residue is purified by normal phase silica gel chromatography (0-20% gradient of MeOH/DCM) to provide 4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-3-nitro-5-(prop-1-ene-2-yl)benzamide (111 mg, 0.211 mmol, 40% yield) as an orange solid. The solid is used without further purification. LCMS ( m/z ): 529.2 [M+H] ⁺ .

Step 4: 3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-isopropylbenzamide

To a solution of 4-((4-((4-carbamoyl-2-nitrophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-3-nitro-5-(prop-1-en-2-yl)benzamide (121 mg, 0.229 mmol) in methanol (20 mL) under nitrogen was added 10% Pd/C (26 mg, 0.024 mmol). The atmosphere of the vessel was exchanged to hydrogen (balloon) and the mixture was stirred overnight. After removal of hydrogen, LCMS analysis showed that the olefin was not completely reduced. The Pd catalyst was removed by filtration and the mixture was concentrated. The residue was then dissolved in methanol (20 mL) and further reduced (H-Cube, 50 psi hydrogen, 30 °C, 1 h, Pd/C cartridge). After evaporation of the solvent, a light brown solid of low purity (~40% by UV) (72 mg) was obtained and used without further purification. LCMS ( m/z ): 471.1 [M + H] ⁺ .

Step 5: 1-(4-(5-Carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-isopropyl-1H-benzo[d]imidazol-5-carboxamide

To a solution of 3-amino-4-((4-((2-amino-4-carbamoylphenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-isopropylbenzamide (72 mg, ~40% purity) in DMF (1.5 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate solution (1 M in 1,4-dioxane, 0.306 mL, 0.306 mmol) at room temperature. The reaction mixture was then stirred at room temperature for 1 h. EDC (110 mg, 0.574 mmol) and TEA (0.160 mL, 1.148 mmol) were then added to the mixture at room temperature. After stirring overnight, the mixture was filtered and the filtrate was directly purified by mass-directed reverse phase HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 30-85% gradient of MeCN/water with 0.1% TFA modifier) to provide 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-isopropyl-1H-benzo[d]imidazole-5-carboxamide (31 mg, 0.039 mmol, 26% yield). LCMS ( m/z ): 793.4 [M+H] ⁺ .

Step 6: (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-isopropyl-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a mixture of 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-isopropyl-1H-benzo[d]imidazole-5-carboxamide (22 mg, 0.028 mmol) and cesium carbonate (46 mg, 0.141 mmol) in DMF (1 mL) was added iodomethane (5 µL, 0.080 mmol) at room temperature. The reaction mixture was then stirred for 5 h. The mixture was filtered and the filtrate was directly purified by mass-directed reverse phase HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of MeCN/water with 0.075% _NH4OH , 10 mM ammonium bicarbonate, pH 10) to afford (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-isopropyl-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (6 mg, 7.3 g) as a white solid. umol, 26% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.14 - 8.05 (m, 3 H), 7.97 - 7.85 (m, 3 H), 7.66 - 7.57 (m, 1 H), 7.54 - 7.43 (m, 2 H), 6.52 (s, 2H), 5.33 - 5.12 (m, 4 H), 4.55 - 4.45 (m, 4 H), 3.61 (s, 3 H), 3.56 (s, 3 H), 2.13 (s, 3 H), 2.12 (s, 3 H), 1.30 - 1.19 (m, 13 H). LCMS ( m/z ): 821.4 [M + H] ⁺ .

Embodiment 38

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-(4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide

To a suspension of 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (1.2 g, 3.56 mmol) and 2,2,3,3-tetrafluorobutane-1,4-diamine, 2 hydrochloride (1 g, 4.29 mmol) in 1-butanol (40 mL) was added sodium bicarbonate (1.078 g, 12.83 mmol) at room temperature. The reaction mixture was then warmed to 120 °C and stirred for 5 days. The mixture was cooled to room temperature and quenched with water. The aqueous phase was extracted with EtOAc (3x). The resulting emulsion was filtered through a cake of Celite. The combined organic layers were washed with brine, dried and concentrated onto Celite. Normal phase chromatography (40 g column, 0-8% gradient of MeOH/DCM) provided 4-((4-amino-2,2,3,3-tetrafluorobutyl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (349 mg, 0.758 mmol, 21.3% yield) as an orange solid. LCMS ( m/z ): 461.2 [M+H] ⁺ .

Step 2: 3-((4-methoxybenzyl)oxy)-5-nitro-4-((2,2,3,3-tetrafluoro-4-((2-nitrophenyl)amino)butyl)amino)benzamide

Sodium bicarbonate (191 mg, 2.27 mmol) was added to a suspension of 1-fluoro-2-nitrobenzene (0.16 mL, 1.52 mmol) and 1-fluoro-2-nitrobenzene (0.16 mL, 1.52 mmol) in 1-butanol (4 mL) at room temperature. Then, the reaction mixture was warmed to 80 ° C and stirred for 12 days. The mixture was cooled to room temperature and quenched with water. The aqueous layer was extracted with EtOAc (3x). The combined organic layer was washed with brine, dried and concentrated. The residue was suspended in DCM. Then, the solid was filtered, washed with DCM and dried to obtain 3-((4-methoxybenzyl)oxy)-5-nitro-4-((2,2,3,3-tetrafluoro-4-((2-nitrophenyl)amino)butyl)amino)benzamide (330 mg, 0.569 mmol, 75% yield) as an orange solid. LCMS ( m/z ): 582.2 [M + H] ⁺ .

Step 3: 3-amino-4-((4-((2-aminophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-((4-methoxybenzyl)oxy)benzamide

To a solution of 3-((4-methoxybenzyl)oxy)-5-nitro-4-((2,2,3,3-tetrafluoro-4-((2-nitrophenyl)amino)butyl)amino)benzamide (328 mg, 0.564 mmol) and ammonium chloride (302 mg, 5.64 mmol) in methanol (5 mL) was added zinc (369 mg, 5.64 mmol) at room temperature. The reaction mixture was then stirred overnight at room temperature. The mixture was filtered through diatomaceous earth, concentrated, and partitioned between water and EtOAc. The aqueous layer was separated and extracted with EtOAc (1x). The combined organic layers were then washed with brine, dried over magnesium sulfate and concentrated. The residue was suspended in DCM. The solid was filtered, washed with DCM and dried to provide impure title compound (70 mg, ~24% yield). The filtrate was concentrated and the residue was purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of MeCN/water with 0.075% NH4OH, 10 mM ammonium bicarbonate, pH 10) to afford pure 3-amino-4-((4-((2-aminophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-((4-methoxybenzyl)oxy)benzamide (92 mg, 0.18 mmol, 31% yield) as a light brown solid. LCMS ( m/z ): 522.3 [M+H] ⁺ .

Step 4: 2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-5-carboxamide

To a solution of 3-amino-4-((4-((2-aminophenyl)amino)-2,2,3,3-tetrafluorobutyl)amino)-5-((4-methoxybenzyl)oxy)benzamide (92 mg, 0.176 mmol) in DMF (1 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate solution (1 M in 1,4-dioxane, 0.370 mL, 0.370 mmol) at 0 °C. Then, the reaction mixture was stirred at room temperature for 1 h. Then, EDC (127 mg, 0.662 mmol) and TEA (0.184 mL, 1.323 mmol) were added to the reaction mixture at room temperature. Then, the reaction mixture was stirred for 2 h. The mixture was diluted with water. The solid was filtered, washed with DCM and water and dried to afford 2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (85 mg, 0.10 mmol, 57% yield) as an off-white solid. LCMS ( m/z ): 844.4 [M+H] ⁺ .

Step 5: (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-(4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a mixture of 2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (65 mg, 0.077 mmol) and cesium carbonate (125 mg, 0.385 mmol) in DMF (1 mL) was added iodomethane (0.012 mL, 0.19 mmol) at room temperature. The reaction mixture was then stirred overnight. The reaction mixture was filtered and the filtrate was directly purified by mass directed reverse phase HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 50-99% gradient of MeCN/water with 0.075% _NH4OH , 10 mM ammonium bicarbonate, pH 10) to afford (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-(4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,2,3,3-tetrafluorobutyl)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (33 mg, 4% by weight) as an off-white solid. 0.038 mmol, 49% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δppm 8.10 (s, 1 H), 7.78 (d, J =1.0 Hz, 1 H), 7.68 - 7.65 (m, 1 H), 7.61 (dd, J =1.9, 7.0 Hz, 1 H), 7.52 (s, 1 H), 7.48 - 7.33 (m, 5 H), 6.77 (d, J =8.6 Hz, 2H), 6.49 (d, J =0.8 Hz, 2 H), 5.25 - 5.09 (m, 4 H), 4.72 (t, J =16.6 Hz, 2 H), 4.49 (q, J =7.0 Hz, 4 H), 3.58 (s, 6 H), 3.49 (s, 3 H), 2.12 (s, 3 H), 2.11 (s, 3 H), 1.23 (t, J =7.1 Hz, 3 H), 1.24 (t, J =7.1 Hz, 3 H). LCMS ( m/z ): 872.4[M + H] ⁺ .

Embodiment 39

(E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-((tetrahydrofuran-3-yl)methoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a solution of (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (49 mg, 0.063 mmol) in DMF (1 mL) was added 3-(bromomethyl)tetrahydrofuran (20.95 mg, 0.127 mmol) followed by potassium carbonate (11.40 mg, 0.083 mmol). The reaction mixture was stirred at 90 °C for 24 h. The mixture was directly purified by preparative HPLC (Phenomenex Eclipse, 5 um packing, 50x30 mm column, 25-55% gradient of MeCN/water with 0.1% TFA modifier). The corresponding fractions were combined and concentrated in vacuo. The residue was partitioned between EtOAc and aqueous sodium bicarbonate solution. The organic layer was separated, dried over sodium sulfate and evaporated in vacuo to afford (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-((tetrahydrofuran-3-yl)methoxy)-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (22.5 mg, 0.027 mmol, 42.6% yield) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.11 - 8.15 (m, 1 H), 8.05 - 8.11 (m, 1 H), 7.99 - 8.05 (m, 1H), 7.79 (s, 2 H), 7.50 (br. s., 3 H), 7.18 - 7.28 ( 3 .59 (s, 3 H), 3.57 (s, 3 H), 3.45 - 3.53 (m, 2 H), 2.11 (s, 3 H), 2.08 (s, 3 H), 1.62 (br. s., 4 H), 1.48 (br. S., 4 H), 1.15 - 1.36 (m, 8 H). LCMS ( m/z ): 833.5 [M + H] ⁺ .

Embodiment 40

(E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a solution of (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (400 mg, 0.484 mmol) in DCM (8 mL) was added HCl solution (4M in dioxane, 0.726 mL, 2.91 mmol) dropwise. Most of the DCM was removed in vacuo. 1,4-Dioxane (8 mL) and more HCl solution (4M in dioxane, 0.726 mL, 2.91 mmol) were added. The vessel was sonicated and stirred vigorously for 2.5 h (gum was still present and the reaction was not complete). The mixture was concentrated again in vacuo and resuspended in THF (6 mL) and water (1 mL). More HCl solution (4M in dioxane, 0.726 mL, 2.91 mmol) was added and the mixture was stirred for 3 h. The reaction mixture was concentrated and dissolved in 20% methanol/DCM (6 mL). To this homogeneous solution was added HCl solution (4M in dioxane, 0.726 mL, 2.91 mmol) and the mixture was stirred for 30 min. Due to the formation of by-products, the solvent was evaporated again and replaced with a 3:1 DCM:ethanol mixture (8 mL) and more HCl solution (4M in dioxane, 0.726 mL, 2.91 mmol). The reaction mixture was stirred at room temperature for 48 h. The mixture was concentrated and distributed between 10% methanol/DCM and saturated sodium bicarbonate aqueous solution. The aqueous layer was extracted twice with 3:1 CHCl ₃ :EtOH. The combined organic phase was concentrated. The residue was partially purified by silica gel chromatography (12 g silica gel; 10-90% [3:1 EA:EtOH]/heptane). The crude solid was suspended in several milliliters of DMSO. The undissolved solid was filtered, washed with DCM, dried and found to be pure enough (~90%, 149 mg) for the synthesis of other analogs. The dissolved material was then purified by mass-directed HPLC (XSELECTCSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of MeCN/water with 0.1% TFA modifier). A few drops of saturated sodium bicarbonate solution were added to each clean fraction. ACN was removed using a stream of nitrogen. The suspended solid was filtered, rinsed with water and dried to provide pure (E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (11.2 mg, 0.016 mmol, 3.2%) as a white solid. ¹ H NMR (DMSO-d ₆ ) δ ppm 8.10 (s, 1H), 7.99 (br. s., 1 H), 7.78 (dd, J =8.4, 1.4 Hz, 1 H), 7.42 (br. s., 1 H), 7.20 (d, J =8.5 Hz, 1 H), 7.12 (t, J =8.0 Hz, 1 H), 7.01 (d, J =7.8 Hz, 1 H), 6.75 (d, J =8.0 Hz, 1 H), 6.48 (s, 1 H 6.32 (s, 1 H), 5.08 (s, 2 H), 4.82 (s, 2 H), 4.44-4.57 (m, 4 H), 3.58 (s, 3H), 3.51 (s, 3 H), 2.14 (s, 3 H), 2.07 (s, 3 H), 1.65 (s, 3 H), 1.48 (s, 3 H), 1.30 (t, J =7.2 Hz, 3 H), 1.25 (t, J =7.2 Hz, 3 H). LCMS ( m/z ): 706.4 [M + H] ⁺ .

Embodiment 41

(E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: 4-(((2S,3S)-4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-diethoxybutyl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide

To a mixture of 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (1.55 g, 4.60 mmol) in 1-butanol (15 mL) was added (2S,3S)-2,3-diethoxybutane-1,4-diamine (1.01 g, 5.75 mmol) and DIEA (2.41 mL, 13.8 mmol). The mixture was stirred at 120°C for 2 h. Then, 4-fluoro-3-nitrobenzamide (0.848 g, 4.60 mmol) was added. The mixture was stirred at 120°C for 18 h. The mixture was cooled and filtered to remove suspended solids. After removal of solvent, silica gel chromatography (40 g silica gel, gradient 5-20% MeOH/DCM) afforded 4-(((2S,3S)-4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-diethoxybutyl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (0.86 g, -25% yield, contaminated with -20% symmetric bis-PMB-protected byproduct) as an orange solid. LCMS ( m/z ): 641.2 [M+H] ⁺ .

The collected precipitate provided 4,4'-(((2S,3S)-2,3-diethoxybutane-1,4-diyl)bis(azanediyl))bis(3-((4-methoxybenzyl)oxy)-5-nitrobenzamide) (423 mg, 0.545 mmol, 12% yield, LCMS ( m/z ): 777.5 [M+H] ⁺ ), which was used to prepare other examples.

Step 2: 3-amino-4-(((2S,3S)-4-((2-amino-4-carbamoylphenyl)amino)-2,3-diethoxybutyl)amino)-5-((4-methoxybenzyl)oxy)benzamide

To a 100 mL round bottom flask was added 4-(((2S,3S)-4-((4-carbamoyl-2-nitrophenyl)amino)-2,3-diethoxybutyl)amino)-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (0.86 g, 1.342 mmol) and methanol (20 mL). To this mixture was added 10 mL of saturated aqueous ammonium chloride solution. To this mixture was added zinc (0.878 g, 13.42 mmol) and the heterogeneous mixture was stirred at room temperature for 15 min. The mixture was filtered and the filter cake was rinsed with MeOH. The filtrate was concentrated.

The crude product was purified by silica gel chromatography (24 g silica gel, 6-20% gradient of MeOH/DCM with 1% _NH4OH as modifier) to afford 3-amino-4-(((2S,3S)-4-((2-amino-4-carbamoylphenyl)amino)-2,3-diethoxybutyl)amino)-5-((4-methoxybenzyl)oxy)benzamide (0.127 g, 16%) as a light yellow solid. LCMS ( m/z ): 581.3 [M+H] ⁺ .

Step 3: 1-((2S,3S)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-5-carboxamide

To a solution of 3-amino-4-(((2S,3S)-4-((2-amino-4-carbamoylphenyl)amino)-2,3-diethoxybutyl)amino)-5-((4-methoxybenzyl)oxy)benzamide (0.127 g, 0.219 mmol) in DMF (6 mL) was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~0.4 in dioxane, 1.094 mL, 0.437 mmol). The mixture was stirred for 15 min. EDC (0.105 g, 0.547 mmol) and TEA (0.152 mL, 1.094 mmol) were added and the reaction was stirred at 50 °C for 18 h. The reaction mixture was poured into 3:1 water:saturated aqueous ammonium chloride solution (20 mL). The resulting solid was filtered, washed with water, and dried to provide 1-((2S,3S)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (0.142 g, 0.157 mmol, 72% yield) as a solid. LCMS ( m/z ): 903.3 [M+H] ⁺ .

Step 4: (E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

To a solution of 1-((2S,3S)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (0.112 g, 0.124 mmol) in DMF (5 mL) was added cesium carbonate (0.121 g, 0.372 mmol) and iodomethane (0.031 mL, 0.496 mmol). The reaction mixture was stirred at room temperature for 18 h. The reaction mixture was diluted with water and extracted with EtOAc (3 x 50 mL). The organic phase was washed with brine (10 mL), dried over magnesium sulfate, filtered and concentrated. The crude product was purified using mass-directed reverse phase HPLC (XSELECT CSHC18, 5 um packing, 150x30 mm column, 15-55% gradient of MeCN/water with 0.075% NH4OH, 10 mM ammonium bicarbonate, pH 10). Pure fractions were combined and concentrated to afford (E)-1-((2S,3S)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-diethoxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-((4-methoxybenzyl)oxy)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (34 mg, 0.36 mmol, 29% yield) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 8.00 - 8.25 (m, 3 H) 7.86 (dd, J =8.49, 1.39 Hz, 1 H) 7.77 (d, J =1.01 Hz, 1 H) 7.62 (s, 1 H) 7.36 - 7.53 (m, 5 H) 6. 75 (d, J =8.62 Hz, 2 H) 6.49 (d, J =6.84 Hz, 2 H) 5.22 (d, J =10.39Hz, 1 H) 5.11 (d, J =10.65 Hz, 1 H) 4.44 - 4.69 (m, 5 H) 4.29 - 4.42 (m, 1 H)3.93 - 4.06 (m, 1 H) 3.63 - 3.84 (m, 3 H) 3.58 (s, 3 H) 3.58 (s, 3 H) 3.40 (s, 3 H) 3.09 - 3.28 (m, 2 H) 2.80 - 2.94 (m, 2 H) 2.12 (s, 3 H) 2 .10 (s, 3H) 1.30 (t, J =7.10 Hz, 6 H) 0.58 (t, J =6.97 Hz, 3 H) 0.47 (t, J =6.97 Hz, 3H). LCMS ( m/z ): 931.4 [M + H] ⁺ .

Table 2 shows Examples 43-92, which can be prepared according to the method exemplified below:

。

.

Embodiment 90

(E)-1-((E)-4-((E)-7-Bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

Step 1: (E)-3-((4-((2-bromo-6-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-4-nitrobenzamide

To a bright yellow suspension of (E)-4-((4-amino-2,3-dimethylbut-2-en-1-yl)amino)-3-nitrobenzamide, hydrochloride (1.05 g, 3.34 mmol) and 1-bromo-2-fluoro-3-nitrobenzene (0.734 g, 3.34 mmol) in 1-butanol (25 mL) was added DIEA (1.75 mL, 10.01 mmol). The reaction was stirred at 80 °C for 5 h and then at room temperature for 16 h. The solid was filtered and rinsed with butanol (20 mL) and water (3 x 20 mL). The solid was dried to provide the title compound (1.16 g, 2.40 mmol, 72% yield) as an orange solid. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.66 (d, J =2.0 Hz, 1 H), 8.3-8.4 (m, 1 H), 7.99 (br s, 1 H), 7.9-8.0 (m, 1 H), 7.87 (dd, J =1.5, 7.8 Hz, 1 H), 7.81 (dd , J =1.5, 8.3 Hz, 1 H), 7.31 (br s, 1H), 6.8-6.9 (m, 1 H), 6.77 (d, J =9.0 Hz, 1 H), 6.06 (t, J =6.0 Hz, 1 H), 3.98 (br d, J =5.8 Hz, 2 H), 3.85 (d, J =6.0 Hz, 2 H), 1.64 (s, 3 H), 1.59 (d, J =1.3Hz, 3 H). LCMS ( m/z ): 478.2 [M + H] ⁺ .

Step 2: (E)-4-amino-3-((4-((2-amino-6-bromophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide

To a mixture of (E)-3-((4-((2-bromo-6-nitrophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)-4-nitrobenzamide (1.06 g, 2.216 mmol) and ammonium chloride (1.778 g, 33.2 mmol) in MeOH (50 mL) cooled in an ice/water bath was added zinc (1.449 g, 22.16 mmol) and the reaction mixture was stirred at room temperature for 40 min. The mixture was filtered through celite and rinsed with methanol. The filtrate was concentrated and purified by silica gel chromatography (24 g silica gel column; 10-60% [3:1 EA:EtOH]/heptane, plus 1% NH ₄ OH solution, 12 min.; 60% gradient of [3:1 EA:EtOH]/heptane, 5 min). The purest fractions were combined and concentrated. The combined fractions were concentrated and purified again (12 g silica gel column; 10-55% [3:1 EA:EtOH]/heptane, no NH ₄ OH modifier, 10 min.; 55% gradient of [3:1 EA:EtOH]/heptane, 5 min.). The fractions were combined and dried to afford the title compound (571 mg, 1.09 mmol, 49.3% yield) as a brown foam. LCMS ( m/z ): 418.3 [M+H] ⁺ , ~80% purity by UV 210-350 nm).

Step 3: (E)-1-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide

To a light brown solution of (E)-4-amino-3-((4-((2-amino-6-bromophenyl)amino)-2,3-dimethylbut-2-en-1-yl)amino)benzamide (570 mg, 1.090 mmol) in DMF (10 mL) cooled in an ice/water bath was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (~1 M in dioxane, 2.18 mL, 2.18 mmol) dropwise rapidly. After stirring for 15 min, EDC (522 mg, 2.73 mmol) and TEA (0.760 mL, 5.45 mmol) were added and the reaction mixture was warmed to room temperature and stirred for 16 h. To the stirred reaction mixture was added a solution of 5:1 water:saturated aqueous NH ₄ Cl (120 mL). The resulting suspension was stirred rapidly for 15 min. The solid was filtered and rinsed with water (3 x 20 mL). The solid was stirred in diethyl ether (15 mL) for 30 min, then filtered and rinsed with diethyl ether. After drying, the title compound (772 mg, 0.94 mmol, 86% yield) was obtained as a light yellow solid. ¹ H NMR(DMSO- d ₆ ) δ: 13.02 (br. s., 1 H), 12.90 (s, 1 H), 8.02 (s, 1 H), 7.94 (br.s., 1 H), 7.71 (d, J =8.4 Hz, 1 H), 7.61 (d, J =7.9 Hz, 1 H), 7. 44 (d, J =7.9Hz, 1 H), 7.35 (br. s., 1 H), 7.26 (d, J =8.4 Hz, 1 H), 7.19 (t, J =8.0 Hz, 1H), 6.64 (s, 1 H), 6.52 (s, 1 H), 5.26 (br. s., 2 H), 5.00 (br. s., 2 H), 4.53-4.63 (m, 4 H), 2.12 (s, 3 H), 2.10 (s, 3 H), 1.69 (br. s., 3 H), 1.63 (br. s., 3 H), 1.35 (t, J =7.1 Hz, 3 H), 1.32 (t, J =7.1 Hz, 3 H). LCMS ( m/z ): 740.2/742.4 [M+H] ⁺ , ~90% purity by UV210-350 nm).

Step 4: (E)-1-((E)-4-((E)-7-bromo-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a solution of (E)-1-(4-(7-bromo-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-1-yl)-2,3-dimethylbut-2-ene-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxamide (760 mg, 0.923 mmol) in DMF (15 mL) was added potassium carbonate (319 mg, 2.309 mmol) and iodomethane (0.13 mL, 2.12 mmol). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was diluted with water (30 mL) and a clumping solid was formed. The mixture was then partitioned between DCM and water. The organic phase was then washed with brine and concentrated. Purification by silica gel chromatography (24 g silica gel column; 10-60% gradient of [3:1 EA:EtOH]/heptane, plus 1% _NH4OH , 15 min.; 60% [3:1 EA:EtOH]/heptane, plus 1% _NH4OH , 10 min.) provided the title compound (386 mg, 0.477 mmol, 51.7% yield) as a light orange foam after evaporation of the solvent. ¹ H NMR(DMSO- d ₆ ) δ: 8.08-8.12 (m, 1 H), 8.01 (br. s., 1 H), 7.80 (dd, J =8.4, 1.4 Hz, 1 H), 7.62-7.67 (m, 1 H), 7.49-7.53 (m, 1 H), 7.43 (br. s ., 1 H), 7.25-7.34(m, 2 H), 6.49 (s, 1 H), 6.44 (s, 1 H), 5.16 (s, 2 H), 4.89 (s, 2 H), 4.47-4.57 (m, 4 H), 3.59 (s, 3 H), 3.56 (s, 3 H), 2. 13(s,3H), 2.12 (s, 3 H), 1.59 (s, 3 H), 1.50 (s, 3 H), 1.24-1.33 (m, 6 H). LCMS ( m/z ): 768.5/770.5 [M +H] ⁺ .

Embodiment 91

(E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-5-carboxamide

Step 1: (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a suspension of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (125 mg, 0.147 mmol, which can be prepared according to the preparation described in Example 14 of PCT International Application (Int. Appl.) WO 2017175147) and potassium carbonate (44.7 mg, 0.324 mmol) in DMF (1.4 mL) was added a solution of iodomethane (0.019 mL, 0.31 mmol) in DMF (0.4 mL). The mixture was stirred at room temperature for 18 h, and then diluted with water. The mixture was extracted with dichloromethane (3X). The combined organic layer was washed with water, dried over sodium sulfate, filtered and concentrated. The residue was purified by reverse phase mass-guided preparative HPLC (XSELECT CSH C18, 5 um filler, 150x30 mm column, 15-55% gradient of ACN/ water, containing 0.1% TFA modifier). The fraction containing the desired product was passed through alkaline PL-HCO3 MP SPE cartridge. The eluent was concentrated to afford (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (45 mg, 0.051 mmol, 34.8% yield). ¹ H NMR (DMSO-d ₆ ) δ ppm 8.04 (br. s., 2H), 7.71-7.75 (m, 2H), 7.38-7.48 (m, 4H), 6.40 (s, 1H), 6.35 (s, 1H), 5.66-5.80 (m, 2H), 4.79-4.89 (m, 4H), 4.38-4.49 (m, 4H), 4.02 (br. t., J=6.3 Hz, 2H), 3.74 (s, 3H), 3.47-3.54 (m, 10H), 2.18-2.29 (m, 6H), 2.12 (s, 3H), 2.10 (s, 3H) , 1.64-1.72 (m, 2H), 1.18-1.25 (m, 6H). LCMS ( m/z ): 878.3 [M + H] ⁺ .

Step 2: (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

To a mixture of (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (26 mg, 0.030 mmol) and NMO (5.20 mg, 0.044 mmol) in tert-butanol (1.2 mL) and water (0.3 mL) was added 2.5% osmium tetroxide in tert-butanol (0.019 mL, 1.5 µmol). The mixture was stirred at room temperature for 64 h and then filtered. The filtrate was directly purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of ACN/water with 0.1% TFA modifier). The fractions containing the title compound were passed through a PL-HCO3 MP SPE cartridge. The eluent was concentrated to afford (E)-1-(4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-7-(3-morpholinopropoxy)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (11 mg, 0.012 mmol, 40.7% yield). 1H NMR (400 MHz, DMSO- d ₆ ) δ ppm 1.24 - 1.29 (m, 6 H) 1.81 - 1.86 (m, 2 H) 2.09 - 2.12 (m, 6 H) 2.27 (br. s., 4 H) 2.35 - 2.39 (m, 2 H) 3.47 -3.53 ( m, 10 H) 3.81 (s, 3 H) 3.82 - 3.93 (m, 2 H) 4.14 (t, J =6.40 Hz, 2 H)4.26 - 4.33 (m, 2 H) 4.44 - 4.58 (m, 6 H) 4.98 (d, J =6.53 Hz, 1 H) 5.07 (d, J =6.02 Hz, 1 H) 6.39 (s, 1 H) 6.44 (s, 1 H) 7.41 - 7.48 (m, 4 H) 7.72 (dd, J =4.52, 1.00 Hz, 2 H) 8.05 (br. s., 2 H). LCMS ( m/z ): 912.2 [M + H] ⁺ .

Embodiment 92

(E)-7-(3-aminopropoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, formate

A mixture of (E)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-hydroxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide (384 mg, 0.513 mmol), tert-butyl (3-bromopropyl)carbamate (733 mg, 3.08 mmol) and _K2CO3 (425 mg, 3.08 mmol) in DMF (10 _mL ) was heated at 90 °C for 9 days. LCMS analysis showed that partial conversion to the O-alkylated intermediate increased over time to reach about 50% conversion in 9 days. The reaction mixture was filtered and concentrated. The residue was purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of ACN/water, containing 0.1% TFA modifier) to provide 100 mg of brown residue after solvent evaporation. The residue was a mixture of NB°C intermediate and the title compound (removal of the B°C group occurred during solvent evaporation). The residue was dissolved in methanol (1 mL) and 1,4-dioxane (2 mL) and sonicated to a brown solution, and then HCl (4 M in dioxane, 1.282 mL, 5.13 mmol) was added. The mixture was sonicated again and stirred at room temperature for 3 h. The reaction mixture was concentrated, dissolved in DMSO, and then purified by mass-directed preparative HPLC (XSELECT CSH C18, 5 um packing, 150x30 mm column, 15-55% gradient of ACN/water with 0.1% formic acid modifier) to afford (E)-7-(3-aminopropoxy)-1-((E)-4-((E)-5-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-2,3-dimethylbut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide, formate salt as a white solid (8.0 mg, 9.4 µmol, 1.8% yield). ¹ H NMR (400MHz, DMSO- d ₆ ) δ ppm 8.36 (s, 1 H), 8.12 (s, 2 H), 8.05 (br. s., 1 H), 7.76-7.84 (m, 2 H), 7.53 (s, 1 H), 7.47 (s, 1 H), 7.44 (s, 1 H), 7.25 (d, J =8.5Hz, 1 H), 6.47 (s, 1 H), 6.32 (s, 1 H), 5.06 (s, 2 H), 4.85 (s, 2 H), 4.41-4.57 (m, 4 H), 4.22 (t, J =6.4 Hz, 2 H), 3.59 (s, 3 H), 3.56 (s, 3 H), 2.77(t, J =6.9 Hz, 2 H), 2.12 (s, 3 H), 2.06 (s, 3 H), 1.84 (quint, J =6.5 Hz, 2H), 1.63 (s, 3 H), 1.49 (s, 3 H), 1.29 (t, J =7.1 Hz, 3 H), 1.24 ( t, J =7.1Hz, 3H). LCMS ( m/z ): 806.3 [M + H] ⁺ .

Table 3 shows Examples 93-109, which can be prepared according to the following exemplified method:

。

.

AlexaFluor-488 FRET Assay Ligand

3',6'-Diamino-5-((2-(1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamido)ethyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-disulfonic acid

1-(4-(5-Carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxylic acid dihydrochloride

To 1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxylic acid methyl ester bis trifluoroacetate (400 mg, 0.434 mmol, Example 23 described in PCT Publication No. WO 2017175147) in THF (3.47 mL), MeOH (3.47 mL), and water (1.74 mL) was added 8 M potassium hydroxide (1.09 mL, 8.68 mmol) at room temperature. After stirring overnight, the reaction was concentrated and water was added. The mixture was acidified to pH 4-5 with 7 N aq HCl, and the resulting grey solid was collected by filtration to give the title compound (335 mg, 0.423 mmol, 97 % yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ ppm 12.82 - 12.95 (m, 3 H), 8.08 (s, 1 H), 7.99 (br. s., 2 H), 7.83 (d, J =8.34 Hz, 1 H), 7.78 (d, J =8.34 Hz, 1 H), 7.5 8 (t, J =7.33 Hz, 2 H), 7.36 (br. s., 1 H), 6.60 (d, J =4.80 Hz, 2 H), 4.58 (d, J =6.57Hz, 4 H), 4.29 (br. s., 4 H,) 2.10 (s, 6 H), 1.88 (br. s., 4 H) , 1.31 (t, J =6.95 Hz, 6 H); LCMS: Rt = 0.83 min, [M+H] ⁺ = 680.5.

Step 1: N- (2-aminoethyl)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide trifluoroacetate

1-(4-(5-Carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazole-5-carboxylic acid (10 mg, 0.015 mmol) was dissolved (with sonication) in DMSO (300 μL) at 37° C. To this was added a solution of (9H-fluoren-9-yl)methyl (2-aminoethyl) carbamate hydrochloride (6.9 mg, 0.022 mmol) and HATU (7.6 mg, 0.020 mmol) in DMSO (100 μL), followed by DIEA (10 μL, 0.057 mmol). After stirring overnight, the reaction was diluted with DMF (600 µL), 4-methylpiperidine (400 µL) was added, and the reaction was stirred at room temperature for 1 h. The mixture was concentrated, and the residue was diluted with 1:1 DMSO: MeOH (<1 mL) and purified by reverse phase chromatography (Jupiter C18 preparative column, 10 mL/min) eluting with 30 - 100 % (9:1 ACN: water) in water (0.1 % TFA additive) to give the title compound (8.45 mg, 10.1 µmol, 69 % yield). LCMS: Rt = 0.62 min, [M+H] ⁺ = 722.4.

Step 2: 3',6'-diamino-5-((2-(1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamido)ethyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-disulfonic acid

N- (2-aminoethyl)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-1-yl)butyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-5-carboxamide trifluoroacetate (8.45 mg, 10.1 μmol) was dissolved in DMF (200 μl) and added to solid (5,6-) Alexa Fluor 488-ONSu (5.00 mg, 7.92 μmol). The commercial Alexa Fluor 488-ONSu reagent is a mixture of 5- and 6-isomers.

When a solution was produced, DIPEA (2 μL, 0.01 mmol) was added and the mixture was stirred overnight in the absence of light (by vortexing). LCMS showed the formation of earlier and later eluting product peaks with the expected molecular weight ([M+H] 1238.6). The reaction was concentrated, and the residue was dissolved in 1:1 DMSO: MeOH (<1 mL) and purified by reverse phase chromatography (Jupiter C18 preparative column, 10 mL/min) using 15 - 100% (9:1 ACN: water) elution in water (0.1% TFA additive). The earlier eluting positional isomers were obtained in high purity. In contrast, the fractions of the later eluting isomers also contained unreacted starting materials. The fractions containing the impure later eluting isomers were combined and concentrated. This residue was dissolved in 1:1 DMSO: MeOH (<1 mL) and purified by reverse phase chromatography (Waters Symmetry Prep column, 10 mL/min) eluting with 15 - 100 % (9:1 ACN: water) in water (0.1 % TFA additive) to give the title compound (late eluting isomer, 1.94 mg, 1.49 µmol, 19 % yield). LCMS: Rt = 0.69 min, [M+H] ⁺ = 1238.6. Note that the proposed structure of the title compound (5-isomer) was not based on rigorous structural determination but on previous observations that the 5-isomer was generally the later eluting isomer by reverse phase HPLC methods.

Bioassays and Data:

As described above, the compounds of the invention are modulators of STING and can be used to treat diseases mediated by STING. The biological activity of the compounds of the invention can be determined using any suitable assays and tissue and in vivo models for determining the activity of compounds as modulators of STING.

The _pIC50 value for each compound is reported in at least one experiment or as an average of multiple experiments. It is understood that the data described herein can have reasonable variations depending on the specific conditions and procedures used by the person performing the experiments.

Binding assay

(1) SPA

，其中U为未知的值，C1为被10 μM cGAMP完全抑制的平均应答和C2最大应答平均值。使用等式

进行曲线拟合，其中A为最小应答，B为最大应答，C为log₁₀ *XC50，D为斜率，和x为ABASE XE中的 log₁₀化合物浓度[M]。在这样的条件下，阳性对照化合物cGAMP的表观抑制常数为250 nM，为其4-5nM的实际亲和力的约50倍(Zhang X等人, Molecular Cell, 2013, 51:1-10)。 A radioligand binding assay was developed to measure the interaction of the compound of formula (I) and the carboxyl terminal domain (CTD) of STING by competing with 3H-cGAMP (tritium-labeled cyclic guanine monophosphate (2', 5') -adenine monophosphate (3', 5')). See also Li et al. (Nature Chemical Biology, 10, 1043-1048, (2014) ). A protein encoding the sequence of human STING spanning residues 149 to 379 (gene ID 340061) was expressed in bacteria with a carboxyl terminal ^Flag® peptide fused to AviTag™ for biotinylation and a six-histidine tag for affinity purification. The purified STING-Flag-AviTag-6Xhis protein was biotinylated to completion using the enzyme BirA (Beckett D. et al., Protein Science, 1999, 8:921-929 ). The relative potency of compounds of formula (I) was determined by competition in an equilibrium binding reaction with 50 nM biotinylated-STING, 50 nM 3H-cGAMP and 1.25 mg/mL streptavidin-coated scintillation proximity assay beads (PerkinElmer) in phosphate-buffered saline buffer containing 0.02% (w/v) Pluronic F127 and 0.02% (w/v) bovine serum albumin in Greiner white 384-well plates (Cat. No. 784075) pre-stamped with 100-250 nL of compound in neat DMSO. The binding reaction was incubated at room temperature for 60 minutes. Luminescence was measured (ViewLux™) and the following formula was used to express the raw reading as % inhibition:

, where U is the unknown value, C1 is the average response completely inhibited by 10 μM cGAMP and C2 is the average of the maximum responses. Using the equation

Curve fitting was performed, where A is the minimum response, B is the maximum response, C is log ₁₀ *XC50, D is the slope, and x is the log ₁₀ compound concentration [M] in ABASE XE. Under such conditions, the apparent inhibition constant of the positive control compound cGAMP is 250 nM, which is about 50 times its actual affinity of 4-5 nM ( Zhang X et al., Molecular Cell, 2013, 51: 1-10).

Compounds of Examples 1-5 and 7-9, 11-14, 16 and 19 were tested using the SPA assay described above and exhibited _pIC50 values ranging from 7 to above the upper assay value of 7.4.

(2) FRET assay

The binding efficacy of the molecule to the C-terminal domain (CTD) of human STING was determined using a competitive binding assay. In this assay, a STING (149-379) recombinant protein with a C-terminal biotinylated Avi-tag was used. When bound to STING, an Alexa488-labeled orthosteric site probe (see pages 347-350 for the synthesis of FRET assay ligands) receives 490 nm emission from Tb-streptavidin-Avi-STING and measures the increase in fluorescence at 520 nm. Molecules competing for the probe binding site will result in a low 520 nm signal. The assay was run in a Greiner black 384-well plate (catalog number 784076) containing 100 nL of compound in pure DMSO. A solution of 500 pM STING, 500 pM Streptavidin-Lumi4-Tb, and 100 nM Alexa488 probe in phosphate buffered saline containing 0.02% (w/v) Pluronic F127 and 0.02% (w/v) bovine serum albumin was added to the plate using a Combi liquid handler (ThermoFisher). The plate was centrifuged at 500 rpm for 1 minute, incubated at room temperature for 15 minutes, and then the fluorescence emission at 520 nm after 337 nm laser excitation was measured on an Envision plate reader (Perkin-Elmer). pIC ₅₀ values were determined using a standard four-parameter curve fit in ABASE XE as described above.

Examples 1-42, 44-89, and 91-108 were tested using the FRET assay described above and exhibited _pIC50 values ranging from 5.0 to above the upper assay value of 9.9.

For example, the _pIC50 of the FRET assay in the following example is:

。

.

Cell function assay

The function of the compounds of formula (I) can be determined in a cellular assay detecting specific activation of STING and/or inhibition of IFNβ protein secretion.

(1) Functional Assay I (PBMC Antagonist Assay): Inhibition of STING by the compound of formula (I) can be determined by measuring the loss of interferon β secreted by peripheral blood mononuclear cells (PBMCs) stimulated with an EC80 concentration of a STING agonist (Example 167 described in PCT Publication No. WO 2017175147) or 77 nM Bacmam virus (a double-stranded DNA virus) and then treated with different doses of the compound of formula (I). Frozen PBMC cells were thawed and diluted to a final concentration of 6×10 ⁵ cells/mL in culture medium (RPMI-1640 containing 1.5 g/L NaHCO ₃ , 4.5 g/L glucose, 10 mM Hepes and 1 mM sodium pyruvate, 10% FBS). PBMC-cell suspensions were distributed at a density of 15,000 cells per well into 384-well tissue culture plates (Griener 781073) containing 250 nL of compound diluted in DMSO. The PBMC plates were incubated for 30 minutes before the addition of STING agonists. The levels of IFNβ protein secreted into the growth medium were measured using a human IFNβ electrochemiluminescence kit (Meso Scale Diagnostics) with STING agonists following the manufacturer's instructions after incubation for 4 hours at 37°C. The percentage of inhibition was determined relative to a control lacking compound treatment or EC80 of STING agonist and plotted as a function of compound concentration to determine _pIC50 using the standard two-state model of receptor-ligand inhibition.

Examples 1-4, 12, 13, 18-20, 23-25, 27, 29-34, 36, 38, 40, 41, 45, 47, 48, 50, 53, 54, 56-60, 63, 65-67, 69-72, 74, 76-89, 91, and 93-105 were tested using Functional Assay I (PBMC Antagonist Assay) described above. Examples 2, 3, 13, 18-20, 23-25, 27, 29-34, 38, 40, 41, 45, 47, 48, 50, 53, 54, 56-60, 63, 65-67, 69-72, 74, 76-89, 91, and 93-105 exhibited pIC50 values ranging from 4.3 to above the upper assay value of _8.1 .

For example, the _pIC50 of the PBMC antagonist assay in the following example is:

。

.

(2) Functional assay II (PBMC agonist assay): The activation of STING by the compounds of formula I was determined by measuring the level of IFNβ secreted by human peripheral blood mononuclear cells (PBMCs) treated with different doses of the compounds of formula (I). Frozen PBMC cells were thawed, resuspended in culture medium (RPMI-1640 containing 1.5 g/L NaHCO ₃ , 4.5 g/L glucose, 10 mM Hepes and 1 mM sodium pyruvate, 10% FBS, 10 ng/mL lipopolysaccharide) to a final concentration of 3×10 ⁵ cells/mL, and plated at a density of 15,000 cells per well into 384-well tissue culture plates (Griener 781073) containing 250 nL of the compound diluted in DMSO. The level of IFNβ protein secreted into the growth medium was measured after incubation at 37° C. for 4 hours using a human IFNβ electrochemiluminescence kit (Meso Scale Diagnostics) following the manufacturer's instructions. Percent activation was determined relative to control DMSO treatment and plotted as a function of compound concentration to determine the pEC50 using standard models of receptor activation.

Examples 1-4, 12, 13, 18-20, 23-25, 27, 29-34, 36, 38, 40, 41, 45, 47, 48, 50, 53, 54, 56-60, 63, 65-67, 69-72, 74, 76-89, 91, 93 and 94-105 were tested using the Functional Assay II (PBMC Agonist Assay) described above. Examples 1-4, 12, 13, 81 and 88 exhibited pEC50 values ranging from 4.3 to 7.3. All other tested compounds exhibited pEC50s below 4.3.

(3) Functional assay III (HEK WT agonist assay): Activation of STING in cells can be determined using a luciferase reporter gene assay in human embryonic kidney cells (HEK293T) co-transfected with a plasmid (Agilent Technologies) expressing STING and the enzyme firefly luciferase driven by the interferon-stimulated response element promoter (pISRE-Luc). Full-length human STING (gene ID 340061) and full-length human cyclic guanine adenine synthetase (cGAS) (reference sequence NM_138441.2) were cloned into a mammalian cell expression vector containing a cytomegalovirus promoter. Transfection was prepared using a cell suspension with ^Fugene® 6 following the manufacturer's instructions (3:1 ^Fugene® : DNA). Fifty microliters of the transfection suspension were dispensed into the wells of a 384-well plate containing 250 nL of the compound of formula (I). The final well composition contained 20,000 cells/well, 1 ng STING, 20 ng pISRE-Luc and empty vector pcDNA3.1 (Invitrogen) to bring the total DNA concentration to 125 ng. Control wells expected to generate maximum activation of STING were co-transfected with cGAS expression plasmid. The plates were sealed and incubated at 37°C for 24 hours. Expression of firefly luciferase was processed using the Steady- ^Glo® luciferase assay system (Promega) and analyzed using a standard laboratory luminescence plate reader. The data were normalized for the luminescence response in the presence of cGAS, plotted as a function of compound concentration, and fitted using a standard model of receptor activation to derive pEC ₅₀ .

Examples 1-9, 11-22, 24-28, 35, 36, 39, 45-52, and 72-74 were tested using the functional assay III (HEK WT agonist assay) described above. Examples 1-9 and 11-16, 24, 26-28, 35, 39, 48, 49, and 73 exhibited pEC ₅₀ values ranging from 5.1 to above the upper assay value of 8.1. The maximum response varied from 5 to 139% of the control wells.

(4) Functional assay IV (THP-1 antagonist assay)

The inhibition of STING by the compounds of formula (I) was determined by measuring the loss of interferon β secreted by human PBMCs or immortalized THP-1 cells stimulated with dsDNA-containing baculovirus (Bacmam virus). THP-1 cells plated at a density of 1×10 ⁵ cells/well in a round-bottom 96-well plate in culture medium (RPMI-1640, which contains 1.5 g/L NaHCO ₃ , 4.5 g/L glucose, 10 mM Hepes and 1 mM sodium pyruvate, 10% FBS, 1% PSF, 50 uM β-MeOH) were incubated with different concentrations of STING antagonists for 60 minutes before adding Bacmam virus (final MOI of 40 pfu/cell). The level of IFNβ protein secreted into the growth medium was measured after 6 and 20 hours of incubation at 37°C using a human IFNβ electrochemiluminescence kit (Meso Scale Diagnostics) following the manufacturer's instructions. IFNβ (pg/ml) levels were converted to percent inhibition relative to controls lacking compound treatment (Control 1) or infected with Bacmam virus (Control 2) and fitted using a sigmoid four-parameter least squares fitting model to define the reported compound potency as pIC50. 100 x (1-(sample well-(Control 2)/(Control 1-Control 2).

Examples 1, 3-5, 7-9, 12-14, 16-20, 22, 24-26, 27, 29, 30, 35-41, 44, 45-56, 59, 60, and 62-77 were tested using the Functional Assay IV (THP-1 Antagonist Assay) described above. Examples 1, 3, 4, 7-9, 12, 13, 16-20, 22, 24-27, 29, 30, 35, 37-39, 41, 44-56, 60, 62-75, and 77 exhibited _pIC50 values ranging from 4.3 to above the upper assay value of 9.1.

For example, the _pIC50 of the THP-1 antagonist of the following example is determined to be:

。

.

Claims (15)

1. A compound or a tautomer or a salt thereof, which has a structure of formula I-2,

in R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently C ₁ -C ₃ alkyl; R ^A1 and R ^A2 are independently H, hydroxyl, COOH or optionally substituted (C ₁ -C ₆ alkyl)oxy-, wherein the optionally substituted (C ₁ -C ₆ alkyl)oxy-alkyl group is optionally substituted with 1-4 substituents each independently selected from the group consisting of hydroxyl, -CO ₂ (R ^f ), -N(R ^e )(R ^f ), optionally substituted phenyl and optionally substituted 5-6 membered heterocycloalkyl, Wherein the optionally substituted phenyl or 5-6 membered heterocycloalkyl is optionally substituted by 1-4 substituents independently selected from the following substituents: (C ₁ -C ₄ alkyl)oxy- and C ₁ -C ₄ alkyl; R ³ and R ⁵ are each independently -CO-N(R ^d )(R ^f ), each of R ^d , R ^e and R ^f is independently H or C ₁ -C ₃ alkyl; B is a substituted -C ₁ -C ₄ alkyl- or a substituted -C ₂ -C ₄ alkenyl-, wherein the alkyl part of the substituted C ₁ -C ₄ alkyl- or substituted -C ₂ -C ₄ alkenyl- is substituted by 1-4 substituents independently selected from the following substituents: halogen, hydroxyl, (C ₁ -C ₄ alkyl)oxy- and C _1-4 alkyl, At least one of R ^x or R ^y is independently C ₁ -C ₄ alkyl and the other is H, or both R ^x and R ^y are independently C ₁ -C ₄ alkyl; or its tautomer, or its salt. 2. The compound or tautomer or salt thereof according to claim 1, wherein R ¹⁴ and R ^c2 are ethyl and R ¹⁵ and R ¹⁷ are methyl. 3. The compound or tautomer or salt thereof according to claim 1, wherein R ^A1 and R ^A2 are each independently optionally substituted (C ₁ -C ₄ alkyl) oxy-, wherein optionally A substituted (C ₁ -C ₄ alkyl)oxy-alkyl group is optionally substituted with 1-2 hydroxy substituents. 4. The compound or tautomer or salt thereof according to claim 1, wherein ^R3 and ^R5 are -CO- _NH2 . 5. The compound or its tautomer or salt according to claim 1, wherein B is _-CH2 - _CH2- substituted by 1-2 hydroxyl substituents. 6. The compound or tautomer or salt thereof according to claim 1, wherein R ¹⁴ , R ¹⁵ , R ^c2 and R ¹⁷ are independently methyl or ethyl; One of R ^A1 and R ^A2 is H and the other of R ^A1 and R ^A2 is optionally substituted (C ₁ -C ₄ alkyl)oxy-, wherein the optionally substituted (C ₁ -C ₄ alkyl)oxy-alkyl group is optionally substituted by 1-2 hydroxyl substituents; Both R ³ and R ⁵ are -CO-NH ₂ ; and B is substituted -CH ₂ -CH ₂ - or substituted -CH=CH-, wherein the substituted -CH ₂ -CH ₂ - or substituted -CH=CH- is substituted by 1-4 substituents each independently selected from the following: hydroxyl and C _1-2 alkyl; and At least one of R ^x or R ^y is independently C ₁ -C ₄ alkyl and the other is H, Or both R ^x and R ^y are independently C ₁ -C ₄ alkyl. 7. The compound or tautomer or salt thereof according to claim 1, which is (E)-1-((2R,3R)-4-((E)-5-aminomethyl with the following structure Acyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-di Hydrogen-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) Amino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

8. The compound or tautomer or salt thereof according to claim 1, which is (E)-1-((2S,3S)-4-((E)-5-aminomethyl Acyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-7-(3-hydroxypropoxy)-3-methyl-2,3-di Hydrogen-1H-benzo[d]imidazol-1-yl)-2,3-dihydroxybutyl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) Amino)-7-methoxy-3-methyl-2,3-dihydro-1H-benzo[d]imidazole-5-carboxamide

9. The compound according to any one of claims 1 to 8, or a tautomer or salt thereof, wherein the salt is a pharmaceutically acceptable salt of the compound. 10. A pharmaceutical composition comprising the compound according to claim 9 or a tautomer or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. 11. Use of the compound according to claim 9 or a tautomer or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease or disease mediated by STING. 12. Use according to claim 11, wherein the disease or condition is selected from the group consisting of systemic lupus erythematosus (SLE), cutaneous lupus, lupus nephritis, psoriasis, diabetes including insulin-dependent diabetes mellitus (IDDM), dermatomyositis , systemic sclerosis (scleroderma) and Sjogren's syndrome (SS), rheumatoid arthritis, psoriatic arthritis, STING-associated vasculitis of infancy onset (SAVI), Aicardi Goutieres syndrome (AGS), chilblains Lupus and mixed connective tissue disease. 13. The use according to claim 11, wherein the disease or condition is chronic lung disease, pulmonary fibrosis or asthma. 14. The use according to claim 11, wherein the disease or condition is Alzheimer's disease, amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTD), multiple sclerosis, Parkinson's disease disease and Huntington's disease. 15. The use according to claim 11, wherein the disease or condition is myocardial infarction, heart failure, congenital heart defect, coronary artery disease, hypertension, cardiomyopathy.